CN108728566A - With the relevant SNP marker of millet thousand grain weight properties and its detection primer and application - Google Patents
With the relevant SNP marker of millet thousand grain weight properties and its detection primer and application Download PDFInfo
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- CN108728566A CN108728566A CN201810339798.0A CN201810339798A CN108728566A CN 108728566 A CN108728566 A CN 108728566A CN 201810339798 A CN201810339798 A CN 201810339798A CN 108728566 A CN108728566 A CN 108728566A
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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
- C12Q2600/13—Plant traits
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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
Abstract
This application discloses a kind of and relevant SNP marker of millet thousand grain weight properties and its detection primer and applications.The application and the relevant SNP marker of millet thousand grain weight properties, the SNP marker are located in the bin labels of Article 7 chromosome 32564634bp to 34552998bp, SNP marker and millet mass of 1000 kernel close linkage.The application and the relevant SNP marker of millet thousand grain weight properties, it can be used for the molecular mark of millet thousand grain weight properties, millet mass of 1000 kernel can be predicted by SNP marker detection, early stage identification or screening breeding to realize millet thousand grain weight properties provide scientific basis, have important theory and practice directive significance to the hereditary and selection or improvement process of accelerating millet variety.The application detects the primer pair of SNP marker, and parting, the difference of detection SNP expression can be carried out to millet mass of 1000 kernel.
Description
Technical field
This application involves Millet Breeding technical fields, are marked with the relevant SNP of millet thousand grain weight properties more particularly to a kind of
Note and its detection primer and application.
Background technology
China is the source area of millet and cultivated area maximum, the highest country of yield, yield account for about the whole world in the world
The 80% of total amount.Meanwhile China is also the country that Genetic Resource of Foxtail Millet quantity is most, diversity is most abundant.
Millet mass of 1000 kernel is the weight of 1,000 seed-grains in grams, it is embody seed-grain size and turgor one
Index is important evidence when examining seed-grain quality and the content and field forecast production of species test.Research and control millet thousand
The relevant gene of grain weight Isoquant character and its position functions etc., for instructing millet genetic breeding to have great importance.
Single nucleotide polymorphism (singlenucleotide polymorphism, SNP) is in genomic level by single
DNA sequence polymorphism caused by the variation of nucleotide base, quantity is more in genome, widely distributed, and genetic stability is good.
With the development of gene sequencing technology and the reduction of cost, to Different Individual same gene or genetic fragment carry out direct Sequencing and
Sequence compares, it will be able to determine base with the presence or absence of variation.Therefore, SNP detections are conducive to Genotyping, are suitable for quickly and advise
Modelling screening is unknown or the relationship of known SNP and certain inhereditary feature.
The exploitation of SSR marker is focused primarily upon about the relevant QTL positioning of Quantitative Characters of Foxtail Millet and SNP marker research at present
It utilizes etc. by way of and the development and application for being detected the SNP marker of polymorphism using group's self-mating system degeneracy gene order-checking are ground
Study carefully and not yet has been reported that.Therefore, carry out the exploitation of Quantitative Characters of Foxtail Millet SNP marker, and establish assist-breeding system, for carrying
High millet yield saves breeding cost and is of great significance.
Invention content
The purpose of the application be to provide it is a kind of it is new with the relevant SNP marker of millet thousand grain weight properties and its detection primer and
Using.
To achieve the goals above, the application uses following technical scheme：
The one side of the application discloses a kind of with the relevant SNP marker of millet thousand grain weight properties, which is located at the
In the bin labels of seven articles of chromosome 32564634bp to 34552998bp, also, SNP marker and millet mass of 1000 kernel are close
It is chain.
It should be noted that the application is found that the sequence of one section and millet mass of 1000 kernel close linkage, this section by research
SNP marker in sequence directly affects millet thousand grain weight properties, this section of sequence, that is, Article 7 chromosome 32564634bp to
The bin of 34552998bp is marked.
Further investigation revealed that in the bin labels of Article 7 chromosome 32564634bp to 34552998bp, especially
It is that the base of SNP marker is T or C, with millet mass of 1000 kernel close linkage.
In a kind of realization method of the application, one of SNP marker, the i.e. sites TGW-07-634 are specifically disclosed,
For sequence where the sites TGW-07-634 as shown in Seq ID No.1, the sites TGW-07-634 are sequences shown in Seq ID No.1
In from 5 ' end the 229th bit base.
It should be noted that bin labels and the millet thousand of Article 7 chromosome 32564634bp to 34552998bp
Grain weight close linkage, wherein may include numerous SNP markers, the sites TGW-07-634 are in a kind of realization method of the application
Confirm with the relevant SNP marker of millet thousand grain weight properties, under the guidance of the application, however not excluded that the bin label in also have
Other SNP markers.
Preferably, the application utilizes composite interval mapping with the relevant SNP marker of millet thousand grain weight properties to SNP marker
Method, using 5% global significance level, the LOD value of QTL detections is 4.1414, and phenotypic variation explanation rate is 4.18%.
The another side of the application discloses a kind of primer pair of the SNP marker of detection the application, and the upstream of the primer pair is drawn
Object is sequence shown in Seq ID No.2, and downstream primer is sequence shown in Seq ID No.3；
Seq ID No.2：5'-GATACAGATACGCACCAAAG-3'
Seq ID No.3：5'-GCTAGGCACCACATACGAC-3'.
The another side of the application discloses a kind of kit of the SNP marker of detection the application, which includes this
The primer pair of application.
Preferably, in the kit of the application further include reagent for PCR amplification.
The another side of the application discloses a kind of method of the SNP marker of detection the application, includes drawing using the application
The kit of object pair or the application carries out PCR amplification to millet genomic DNA to be measured, passes through the survey of pcr amplification product
Sequence is as a result, obtain the base situation of SNP marker.
The another side of the application discloses a kind of method of prediction millet mass of 1000 kernel, includes the primer pair using the application,
Or the kit of the application passes through the sequencing of pcr amplification product to predicting that the millet genomic DNA of object carries out PCR amplification
As a result, obtaining the base situation of the SNP marker of the application, the mass of 1000 kernel of millet is predicted according to the base situation of SNP marker.
The SNP marker for disclosing the application on one side again of the application is early in the prediction of millet mass of 1000 kernel, millet thousand grain weight properties
Application in phase identification or millet molecule assisting sifting breeding.
The primer pair for disclosing the application on one side again of the application or the kit of the application are in the prediction of millet mass of 1000 kernel, paddy
Application in sub- thousand grain weight properties early stage identification or millet molecule assisting sifting breeding.
Due to using the technology described above, the advantageous effect of the application is：
The application and the relevant SNP marker of millet thousand grain weight properties, can be used for the molecular labeling of millet thousand grain weight properties
Assistant breeding, by the SNP marker detection can predict millet mass of 1000 kernel, for realize millet thousand grain weight properties early stage identification or
Screening breeding provides scientific basis, has important theory and practice to the hereditary and selection or improvement process of accelerating millet variety
Directive significance.The application detects the primer pair of SNP marker, and parting, the difference of detection SNP expression can be carried out to millet mass of 1000 kernel
It is different.
Description of the drawings
Fig. 1 is millet Parent genome dna electrophoresis figure in the embodiment of the present application, wherein the first swimming lane is M swimming lanes, table
Show that DL2000marker, the second swimming lane are swimming lane expression male parent " the Ji paddy No.1 " genomic DNA for being labeled as 1, third swimming lane
The swimming lane for being labeled as 2 indicates maternal " A2 " genomic DNA；
Fig. 2 is the PCR product electrophoretogram of millet Parent genomic DNA in the embodiment of the present application, wherein the first swimming lane is
The swimming lane of A26-1 is marked, indicates that the second swimming lane is using maternal " A2 " genomic DNA as the PCR product that template, 26_1 are primer
It is labeled as the swimming lane of 326-1, is indicated using male parent " Ji paddy No.1 " genomic DNA as the PCR product that template, 26_1 are primer,
Third swimming lane is M swimming lanes, indicates DL2000marker；
Fig. 3 is the comparison of the sequence in maternal and male parent SNP marker where the sites TGW-07-634 in the embodiment of the present application
Result figure, the sites TGW-07-634 are maternal in the bin labels of Article 7 chromosome 32564634bp to 34552998bp
Base is C, and male parent base is T.
Specific implementation mode
This application provides SNP marker, detection primer and its applications with the relevant bin marked regions of millet mass of 1000 kernel.
Millet mass of 1000 kernel close linkage SNP marker TGW-07-634 is located at Article 7 chromosome 32564634bp to the
In the bin labels of 34552998bp, specifically it is located at 32564634bp.
It should be noted that bin refers in Bin mappings (i.e. Bin map), using whole in mapping population in the application
Recombination site information obtains the least unit (Recombination bin) for constituting recombination event.Using obtained bin as
Label is for subsequent linkage map structure and QTL positioning.
In the application, genetic map (genetic map or linkage map) refers to using recombination fraction between genetic marker as base
The linear array figure of relative position between the label of plinth structure.Based on high throughput weight sequencing technologies by mapping population parent and
Progeny population is sequenced, and using a certain number of continuous SNP as the foundation for judging filial generation recombination site, obtains each filial generation
Full-length genome physics recombinates collection of illustrative plates.Using recombination site information whole in mapping population, the minimum for constituting recombination event is obtained
Unit (Recombination bin) and Bin figures (i.e. Bin map), and the bin utilized is used for subsequent company as label
Lock map construction and QTL positioning.
The application offer millet male parent " Ji paddy No.1 " and maternal " A2 " material, the F1 generation material of the two hybridization, and from
Hand over 441 parts of materials of F2 groups in 13 generations.Each part material is planted in Sanya, Hainan, and records and arrange the characters numbers such as mass of 1000 kernel
According to.After each part material progress degeneracy genome is resurveyed sequence, compares the splicing of reference gene group and complete, obtain SNP polymorphism marks.
By thousand grain weight properties data, associated SNP marker is obtained, and SNP marker is verified by cloning and sequencing.
The application is described in further detail below by specific embodiments and the drawings.Following embodiment is only to the application
It is further described, should not be construed as the limitation to the application.
One high-flux sequence of embodiment and SNP marker information analysis
This example utilizes millet male parent " Ji paddy No.1 " and maternal " A2 " material, the F1 generation material of the two hybridization, and selfing
441 parts of materials of F2 groups in 13 generations, i.e. recombinant inbred lines (Recombinant inbred lines, abridge RILs).By each part
Material is planted in Sanya, Hainan, and records and arrange the trait datas such as mass of 1000 kernel.Each part material progress degeneracy genome is resurveyed
After sequence, compares the splicing of reference gene group and complete, obtain SNP marker.By analyzing thousand grain weight properties data, obtain associated
SNP marker, and SNP marker is verified by cloning and sequencing.
This example utilizes RADseq methods, extracts the genomic DNA of each individual of sample, amounts to 444 samples, including it
In 441 RILs, 2 parents, 1 F1 individual.Quality testing is carried out to the genomic DNA of extraction, enzyme is carried out to qualified DNA
It cuts, electrophoresis recycles DNA fragmentation, and carries out cluster preparations plus connector, finally upper machine sequencing.
This example is as follows：
(1) the fresh blades of 1.0g are weighed, shreds and is put into mortar, with 1.5 × CTAB of 3mL are added after liquid nitrogen grinding, are ground into
Homogenate is transferred in the centrifuge tube of 15mL, and 1.5 × CTAB of 1mL are then added into mortar rinses, then is transferred in centrifuge tube, mixing
Afterwards in 65 DEG C of water-bath 30min, during which slowly shake up frequently.
1.5 × CTAB of wherein 1L is formulated：The Tris.Cl75mL of the pH8.0 of CTAB, 1mol/L of 15g,
EDTA 30mL, the NaCl 61.4g of 0.5mol/L, adds deionized water to be settled to 1L；Using it is preceding be added about 2mL mercaptoethanol,
Make its final concentration of 0.2%.
(2) it after the completion of water-bath, is cooled to room temperature, isometric chloroform/isoamyl alcohol is added, gently mixing, until subnatant becomes
Bottle green；Wherein the volume ratio of chloroform/isoamyl alcohol is chloroform:Isoamyl alcohol is 24:1.
(3) 4200rpm centrifuges 10min, and upper strata aqueous phase is moved on to new 15mL centrifuge tubes, adds 2 times of volumes to be pre-chilled anhydrous
Ethyl alcohol mixes static 5min.30min, which is placed, in -20 DEG C precipitates DNA.
(4) 4200rpm centrifuges 10min, discards supernatant, 75% ethyl alcohol of 1mL washing precipitation is added 1 time, and it is dry to be inverted centrifuge tube
50 μ L TE dissolving DNAs are added in dry DNA.
(5) concentration for detecting DNA, is used in combination water to be adjusted to 20ng/ μ L.
(6) it using PstI enzymes to carry out digestion, interrupts genomic DNA, the endonuclease reaction system of PstI enzymes is 30 μ L, including：
20 μ L of genomic DNA, 1 μ L of PstI enzymes, 10 × PstI buffer solutions, 3 μ L, 6 μ L of Nuclease-free water.Endonuclease reaction body
After system prepares, in 37 DEG C of constant-temperature incubations at least 1 hour.
(7) adjunction head reacts, and sequence measuring joints are added on the genomic DNA fragment interrupted, and the system of adjunction head reaction is
40 μ L, including：30 μ L of PstI digestion products, 1 μ L of sequence measuring joints, 10 × connection buffer 1 μ L, 2 μ L of T4DNA ligase,
rATP 0.4μL、ddH2O 5.6μL。
(8) each sample respectively takes 1 μ L reaction products, is added in a new centrifuge tube, one group, every group of every 12 samples
Total volume be 12 μ L.
(9) 300-700bp size segments are cut after dyeing with 3% recycling gel electrophoresis 1h, EB.It is carried out with QIAquick Kit
Glue purification recycles, and recovery product is dissolved in the EB solution of 30 μ L.
(10) PCR reactions are carried out, PCR amplification, PCR are carried out to the segment for making sure to keep in mind recycling using the universal primer of sequence measuring joints
25 μ L of reaction system, including：10 × buffer, 2.5 μ L, the 0.25 μ L of dNTPs of 25mM, 5U/ μ L 0.25 μ L of high fidelity enzyme,
It is 10 μM of 0.5 μ L of sequence measuring joints forward primer, 10 μM of 0.5 μ L of sequence measuring joints reverse primer, 20 μ L of gel extraction product, sterile
1 μ L of water.
PCR reaction conditions are 94 DEG C of pre-degeneration 5min, are recycled subsequently into 10：94 DEG C are denaturalized 30 seconds, 60 DEG C of annealing 30
Second, 72 DEG C extend 40 seconds, after circulation terminates 72 DEG C extension 3min.
(11) library is built in magnetic beads for purifying, completion.
Specifically purification process is：First, 1.2 times of volume magnetic beads are added into PCR product, stand 10min.Then, it is placed in
It is adsorbed on magnetic frame, removes supernatant.Then, 500 μ L, 70% ethanol wash is added twice.After being evaporated on xeothermic instrument, 15 μ L are added
EB solution dissolves 5min.Finally, it is adsorbed on magnetic frame, transfer supernatant is in 1.5mL centrifuge tubes.
(12) detection reaches upper machine sequencing after machine standard.
As a result：Digestion is carried out to 444 samples and builds library sequencing, obtains 75.99Gb initial data, average each individual
171.54Mb.Sequencing sequence is compared onto reference gene group, average comparison rate 86.326%, average coverage rate 8.226% is put down
Sequencing depth be 3.655 ×.
Wherein, 444 samples include 441 RILs, 2 parents, 1 F1 individual；Reference gene group, that is, the Yugu of this example
Genome can obtain Yugu genome sequences from following Internet address hffp：
https://www.ncbi.nlm.nih.gov/genome/？Term=Setaria+italica+ (foxtail+
millet)。
By sequencing and information analysis, there are 33771 polymorphic SNP markers between acquisition parent.According to the side of window sliding
Method, it is a window to choose several SNP, slides a SNP every time and determines the genotype of each window and each individual
Site is exchanged, bin genotype and bin figures are generated.According to bin genotype datas, with MSTMap software building genetic maps, 2022
A bin is navigated on 9 chromosomes, then genetic map data is imported into MapChart softwares, integrates a genome heredity
Linkage map.
Using constructed millet dense genetic map, (CIM) is analyzed using composite interval mapping, to millet mass of 1000 kernel
Trait phenotypes carry out qtl analysis.QTL detections use 5% global significance level, according to 500 permutation tests as a result, determining thousand
The critical LOD value of grain principal characteristic shape phenotypic data qtl analysis, analysis obtain and the relevant sections prediction bin of mass of 1000 kernel and SNP site
Information.
The results show that being located at genetic linkage maps Article 7 chromosome with the relevant SNP marker of millet thousand grain weight properties
169.31 centimorgans (cM), the 191st bin to the 192nd bin (i.e. chr7_bin191-chr7_bin192), that is, Article 7
In the bin labels of chromosome 32564634bp to 34552998bp；These and the relevant SNP marker of millet thousand grain weight properties
Base be all T or C；Wherein, the sites TGW-07-634 are also in the area, specific positioned at the of Article 7 chromosome sequence
The positions 32564634bp.Using composite interval mapping method, using 5% global significance level, the LOD value of QTL detections is
4.1414, phenotypic variation explanation rate is 4.18%, and additive effect value (Additive effect, A) is -0.131.
Two SNP marker of embodiment is verified
According to the bin of prediction labels and SNP site, reference gene group is compared, relevant range gene order is obtained, chosen
300bp or so before and after SNP site designs and develops SNP marker primer, and PCR expansions are carried out as template using male parent and female parent material DNA
Increase.Selection primer amplification is normal, the amplified production of the compound prediction size of pcr amplification product carries out gel extraction, and is sequenced,
Selection male parent and female parent material amplification gene sequence have the labeled primer of SNP site difference.Using F1 generation and F2 for material as masterplate,
Carry out SNP marker primer PCR amplification and result sequencing, selection meet Genotyping and with the relevant labeled primer of phenotypic character.
First, the sequencing result after being recycled according to PCR product selects male parent and female parent material amplification gene sequence SNP
The label of point difference.It is as follows：
(1) according to step (1) to (4) in embodiment one, male parent and female parent gene group DNA are extracted respectively with CTAB methods.
(2) with 0.8% agarose gel electrophoresis Detection and Extraction male parent and female parent gene group DNA, testing result such as Fig. 1
It is shown, wherein the second swimming lane is swimming lane expression male parent " the Ji paddy No.1 " genomic DNA for being labeled as 1, and third swimming lane marks
Maternal " A2 " genomic DNA is indicated for 2 swimming lane.It can be seen that male parent and female parent gene group DNA are extracted successfully.
(3) by the male parent of extraction and female parent gene group DNA be stored in -20 DEG C it is spare.
(4) it respectively using the male parent of extraction and female parent gene group DNA as template, is combined to base using 26_1F and 26_1R primers
Because a group DNA is expanded.Wherein, 26_1F is sense primer, is sequence shown in Seq ID No.2；26_1R is downstream primer, is
Sequence shown in Seq ID No.3；
Seq ID No.2：5'-GATACAGATACGCACCAAAG-3'
Seq ID No.3：5'-GCTAGGCACCACATACGAC-3'.
25 μ L of PCR reaction systems, including：The Taq enzyme of 10 × buffer, 2.5 μ L, the 0.15 μ L of dNTPs of 25mM, 5U/ μ L
It is 0.15 μ L, 10 μM of 0.5 μ L of 26_1F sense primers, 10 μM of 0.5 μ L of 26_1R downstream primers, 1.0 μ L of genomic DNA, sterile
20.2 μ L of water.
PCR reaction conditions are 94 DEG C of pre-degeneration 5min, are recycled subsequently into 35：94 DEG C are denaturalized 30 seconds, 60 DEG C of annealing 30
Second, 72 DEG C extend 40 seconds, after circulation terminates 72 DEG C extension 3min.
In 4 DEG C of preservations after pcr amplification product is purified.Each pcr amplification product takes part to carry out 1% agarose gel electrophoresis
Detection, the results are shown in Figure 2.In Fig. 2, the first swimming lane is the swimming lane for marking A26-1, and expression is with maternal " A2 " genomic DNA
The pcr amplification product of template, the second swimming lane are the swimming lane for being labeled as 326-1, are indicated with male parent " Ji paddy No.1 " genomic DNA
For the pcr amplification product of template.Fig. 2's the results show that primer 2 6_1F and 26_1R can be to male parents and maternal genomic DNA
It amplifies and meets expected target fragments.
Male parent and maternal genomic DNA amplification product are sequenced respectively, primer 2 6_1F and 26_1R expand to obtain
Male parent amplified production sequencing sequence sequence as shown in Seq ID No.1, the sites TGW-07-634 are shown in Seq ID No.1
In sequence from 5 ' end the 229th bit base.Sequencing sequence comparison result such as Fig. 3 institutes of male parent amplified production and maternal amplified production
Show, wherein the base of background mark ash indicates that the base in the sites TGW-07-634, maternal base are C, and male parent base is T.
Then, according to sample mass of 1000 kernel data screening：Male parent mass of 1000 kernel is 3.3 grams, and maternal mass of 1000 kernel is 2.8 grams.441 parts
In sample, 1.6 grams of minimum value, 3.9 grams of maximum value；In 50 samples of mass of 1000 kernel minimum, sample number identical with maternal SNP is more
In 75%；In maximum 50 samples of mass of 1000 kernel, sample number identical with male parent SNP is more than 75%.
The foregoing is a further detailed description of the present application in conjunction with specific implementation manners, and it cannot be said that this Shen
Specific implementation please is confined to these explanations.For those of ordinary skill in the art to which this application belongs, it is not taking off
Under the premise of conceiving from the application, a number of simple deductions or replacements can also be made.
SEQUENCE LISTING
<110>Academy of Agriculture, Zhangjiakou City's (high and cold crop research institute in Hebei province)
<120>With the relevant SNP marker of millet thousand grain weight properties and its detection primer and application
<130> 17I25628
<160> 3
<170> PatentIn version 3.3
<210> 1
<211> 237
<212> DNA
<213>Sequence where the sites TGW-07-634
<400> 1
tacagatacg caccaaagag atgcatagat ttgctgttgg tggagacgct tctcactctc 60
tctacaaagg tggtcttact gatcctggag aggtaaaact cctgtttctt ctattaaagt 120
aatgtgtgct gggagcccag cccccgagcc tgggagccgg ctgagccaca taagcacgtt 180
gagtcgccac ctgcccaccc taggtcagcc gagccgcggg aggcacactg agtcgta 237
<210> 2
<211> 20
<212> DNA
<213>Artificial sequence
<400> 2
gatacagata cgcaccaaag 20
<210> 3
<211> 19
<212> DNA
<213>Artificial sequence
<400> 3
gctaggcacc acatacgac 19
Claims (10)
1. a kind of and relevant SNP marker of millet thousand grain weight properties, it is characterised in that：The SNP marker is located at Article 7 dyeing
In the bin labels of body 32564634bp to 34552998bp, the SNP marker and millet mass of 1000 kernel close linkage.
2. SNP marker according to claim 1, it is characterised in that：The base of the SNP marker is T or C.
3. SNP marker according to claim 1 or 2, it is characterised in that：The SNP marker includes the sites TGW-07-634,
For sequence where the sites TGW-07-634 as shown in Seq ID No.1, the sites TGW-07-634 are Seq ID No.1
In shown sequence from 5 ' end the 229th bit base.
4. SNP marker according to claim 3, it is characterised in that：SNP marker is used using composite interval mapping method
The LOD value of 5% global significance level, QTL detections is 4.1414, and phenotypic variation explanation rate is 4.18%.
5. a kind of test right requires the primer pair of 1-4 any one of them SNP markers, it is characterised in that：The primer pair
Sense primer is sequence shown in Seq ID No.2, and downstream primer is sequence shown in Seq ID No.3；
Seq ID No.2：5'-GATACAGATACGCACCAAAG-3'
Seq ID No.3：5'-GCTAGGCACCACATACGAC-3'.
6. a kind of test right requires the kit of 1-4 any one of them SNP markers, it is characterised in that：In the kit
Including the primer pair described in claim 5；Preferably, in the kit further include reagent for PCR amplification.
7. a kind of method that test right requires 1-4 any one of them SNP markers, it is characterised in that：Including being wanted using right
The kit described in primer pair or the claim 6 described in 5 is sought, PCR amplification is carried out to millet genomic DNA to be measured, is led to
The sequencing result for crossing pcr amplification product obtains the base situation of the SNP marker.
8. a kind of method of prediction millet mass of 1000 kernel, it is characterised in that：Include the primer pair used described in claim 5, or
Kit described in claim 6 passes through pcr amplification product to predicting that the millet genomic DNA of object carries out PCR amplification
Sequencing result obtains the base situation of claim 1-4 any one of them SNP markers, according to the base feelings of the SNP marker
Condition predicts the mass of 1000 kernel of millet.
9. being reflected in the prediction of millet mass of 1000 kernel, millet thousand grain weight properties early stage according to claim 1-4 any one of them SNP marker
Application in fixed or millet molecule assisting sifting breeding.
10. the kit described in primer pair according to claim 5 or claim 6 is in the prediction of millet mass of 1000 kernel, millet
Application in thousand grain weight properties early stage identification or millet molecule assisting sifting breeding.
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| CN201810339798.0A CN108728566B (en) | 2018-04-16 | 2018-04-16 | SNP (Single nucleotide polymorphism) marker related to thousand grain weight traits of millet as well as detection primer and application thereof |
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103502456A (en) * | 2011-02-28 | 2014-01-08 | 巴斯夫植物科学有限公司 | Plants having enhanced yield-related traits and producing methods thereof |
| CN106939342A (en) * | 2017-04-10 | 2017-07-11 | 山西省农业科学院谷子研究所 | A kind of and the cream-coloured chain SNP marker of millet, primer and application |
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| CN103502456A (en) * | 2011-02-28 | 2014-01-08 | 巴斯夫植物科学有限公司 | Plants having enhanced yield-related traits and producing methods thereof |
| CN106939342A (en) * | 2017-04-10 | 2017-07-11 | 山西省农业科学院谷子研究所 | A kind of and the cream-coloured chain SNP marker of millet, primer and application |
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| KAI ZHANG等: "Identification of QTLs for 14 Agronomically Important Traits in Setaria italica Based on SNPs Generated from High-Throughput Sequencing", 《G3: GENES GENOMES GENETICS》 * |
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