CN109706263A - Chain SNP marker and application with wheat stripe rust resisting ospc gene QYr.sicau-1B-1 - Google Patents
Chain SNP marker and application with wheat stripe rust resisting ospc gene QYr.sicau-1B-1 Download PDFInfo
- Publication number
- CN109706263A CN109706263A CN201910134070.9A CN201910134070A CN109706263A CN 109706263 A CN109706263 A CN 109706263A CN 201910134070 A CN201910134070 A CN 201910134070A CN 109706263 A CN109706263 A CN 109706263A
- Authority
- CN
- China
- Prior art keywords
- wheat
- stripe rust
- rust resisting
- sicau
- qyr
- 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.)
- Granted
Links
Landscapes
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The invention belongs to molecular breeding technology fields, provide the SNP marker and application chain with wheat stripe rust resisting ospc gene QYr.sicau-1B.1, the SNP marker is KASP-Yr, can be obtained by nucleotide sequence primer amplification as shown in NO.1~3 SEQ ID.Detection and analysis show that the molecular labeling can accurately track the wheat stripe rust resisting disease QTL QYr.sicau-1B.1, predict the stripe rust resisting characteristic of wheat, and then facilitate carry out Molecular design breeding.The invention also discloses a kind of methods of molecular labeling for identifying wheat stripe rust resisting disease QTL QYr.sicau-1B.1, it can reinforce the accuracy of stripe rust resisting prediction using method provided by the present invention, have the wheat breed for increasing stripe rust resisting or strain for breeding quickly to filter out, the breeding process of improving yield of wheat amount kind can be greatly speeded up.
Description
Technical field
Molecular biology of the present invention and field of crop genetic breeding, more particularly to relevant to wheat stripe rust resisting characteristic of disease shape
Molecular labeling, and in particular, to wheat stripe rust resisting ospc gene QYr.sicau-1B-1 chain SNP marker expands its
The application of primer pair and the molecular labeling.
Background technique
Wheat is the important cereal crops in China, and long-term cultivated area accounts for about cereal crops face at 20,000,000 hectares or more
Long-pending 27%；Total output is 100,000,000 tons or more, accounts for about the 22% of cereal crops yield.Stripe rust of wheat is in China's Wheat Production
A kind of important disease, when the infringement by strip rust bacteria, wheat leaf blade will appear the necrotic plaque of yellow, and can generate yellow
Uredospore, these spores to later period are changing to black；Strip rust bacteria will cause blade chlorisis, can not carry out photosynthesis, from
And the underproduction, even it will cause that No kernels or seeds are gathered, as in a year of scarcity when serious.Many researchs are all being dedicated to new resistant wheat both at home and abroad all the time
The excavation of the research of kind, resistant gene plays an important role breeding and popularization disease-resistant variety.
Wheat line ' 20828 ' always shows as domestic stripe rust prevalence biological strain in southwestern ecotope nearly more than 10 year
Nearly immune resistance also has the characteristics that more spikelet number, proper height and plant type are compact, is breeding first choice parent.
Single nucleotide polymorphism (Single Nucleotide Polymorphism, SNP) refers to DNA in genome
There is DNA sequence polymorphism caused by the variation such as conversion, transversion, insertion, missing on one specific nucleotide position.Its technology is
Sequence information is known using oneself to compare searching SNP site, and the primer for recycling the variant sites of excavation to design specificity comes to base
Because group DNA or cDNA carries out PCR amplification, the specific polymorphism product based on SNP site is obtained, finally utilizes electrophoretic techniques point
The polymorphism of division object.The advantages of SNP marker is that quantity is more, widely distributed；It is distributed not in individual gene and whole gene group
Uniformly；SNP gene frequency is readily estimated.
KASP be by LGC company (Laboratory of the Government Chemist) (http: //
Www.lgcgenomics.com) competitive ApoE gene technology (the Kompetitive Allele researched and developed
Specific PCR, KASP) have the characteristics that low cost, high-throughput novel gene typing method, pass through prime end base
Special matching to carry out accurately diallele parting to the site SNP and In Del, in rice, wheat, Soybean and Other Crops
Molecular marker assisted selection in be used widely.
There is scholar to carry out QTL positioning to stripe rust resisting character before this, finds associated QTL in wheat extensively
In the presence of, and be distributed in each chromosome, the stripe rust resisting QTL wherein navigated on 1B chromosome is most.But due to disease
The height of opportunistic pathogen microspecies makes a variation, and a large amount of Stripe Rust Resistance Gene or is losing resistance, therefore all the time both at home and abroad
Many researchs are all in the research for being dedicated to new resistant wheat kind, this is just it should be understood that the disease-resistant gene and its something lost in the anti-source of wheat
Biography feature cultivates more efficient resistant variety to carry out more rationally effective application to the anti-source of wheat.
Summary of the invention
The purpose of the present invention is to provide the molecule marks with wheat stripe rust resisting disease QTL QYr.sicau-1B.1 close linkage
Note.
Another object of the present invention is to provide the fluorescence quantification PCR primers of the molecular labeling.
Third object of the present invention is to provide above-mentioned wheat stripe rust resisting disease QTL QYr.sicau-1B.1 close linkage
Molecular labeling application.
The purpose of the present invention is what is be achieved through the following technical solutions:
Based on object above, applicant is female parent using stripe rust resisting wheat strain ' 20828 ', with wheat breed ' CN16 '
For paternal hybrid, hybrid F is obtained1, F1F is obtained for individual plant selfing2, in F2Using single seed descent, until F8In generation, is contained
199 recombinant inbred lines for being constitute genetic mapping group.To recombinant inbred lines stripe rust phenotypic evaluation, parent is extracted
' 20828 ', ' CN16 ' and recombinant inbred lines plant DNA, this research position the anti-item of wheat using wheat 55K SNP chip
Rust QTL.Wheat 55K SNP chip is a economical middle density SNP developed on the basis of wheat 660K SNP chip
Chip.Chip includes 55,000 or so wheat SNP marker, is evenly distributed on 21 chromosomes, is put down on every chromosome
There are 2,600 labels, the average genetic between marking is about 0.1cM, and average physical distance is less than 300Kb, is suitable for one
As Germplasm Resources Diversity analysis, genetic mapping and new gene excavations, the registration of icp gene group analysis, kind (refer to identification
Line analysis).
According to 55K SNP chip data, genetic map is constructed using JoinMap4.0.In conjunction with the stripe rust Phenotype Number of group
According to complete Interval Mapping (the Inclusive Composite Interval in QTL IciMapping 4.0
Mapping-ADD, ICIM-ADD), it is arranged under conditions of threshold values LOD >=2.5, with tri- times of 2016-2018 totally 4 ecosites
And BLUP (optimal linear unbiased prediction, best linear unbiased prediction) value of 4 ecosite stripe rust
QTL is detected, the 1.3cM deciding field on 1B chromosome long arm goes out to stablize the wheat stripe rust resisting disease QTL of expression
QYr.sicau-1B.1 carries out physical positioning to flanking marker and is located at the gene in section every 1Mbp screening, and screening obtains altogether
61 genes are obtained, and these genes are cloned in parent ' 20828 ' and ' CN16 ', to acquisition polymorphic site and are divided
The exploitation of son label, devises 10 pairs of totally 30 KASP primers (table 1) altogether, finally obtains label KASP-Yr and stripe rust resisting
QTL QYr.sicau-1B.1 close linkage.
1 10 pairs of KASP primer sequences of table
Wheat stripe rust resisting disease QTL QYr.sicau-1B.1 of the present invention, from female parent ' 20828 ', which is located at
It is 474Mb in the physical location of RefSeqv1.0 genome version on chromosome of wheat 1B is long-armed.The present invention provides above-mentioned small
Application of the wheat stripe rust resisting QTL QYr.sicau-1B.1 in regulation wheat stripe rust resisting characteristic of disease shape.
Further, the present invention provides the molecular labeling KASP-Yr of wheat stripe rust resisting disease QTL QYr.sicau-1B.1,
Itself and wheat stripe rust resisting disease QTL QYr.sicau-1B.1 close linkage.The molecular labeling and wheat stripe rust resisting disease QTL
QYr.sicau-1B.1 common location is on wheat 1B chromosome long arm, and the genetic distance between QYr.sicau-1B.1 is small
In 1.3cM.The polymorphism of the SNP marker is A/G.Nucleotide sequence containing the SNP marker such as SEQ ID
Shown in NO.31.
The molecular labeling KASP-Yr of wheat stripe rust resisting disease QTL QYr.sicau-1B.1 of the invention is by nucleotide sequence
The primer pair PCR amplification as shown in NO.1~3 SEQ ID obtains.Above-mentioned 3 primers include 2 upstream primers and 1 it is general under
Primer is swum, different fluorophors is modified at 2 upstream primers 5 ' end respectively；The nucleotide sequence of 3 primers is respectively such as
Shown in SEQ ID NO.1,2 and 3.
The SNP marker be averaged LOD value be 21.39, explain about 25.52% phenotypic variation.
The present invention provides for detecting the chain SNP marker of wheat stripe rust resisting disease QTL QYr.sicau-1B.1
Specific primer sets contain nucleotide sequence primer as shown in NO.1~3 SEQ ID.
Kit containing above-mentioned specific primer sets belongs to the scope of protection of the present invention.
The present invention provides above-mentioned molecular labeling KASP-Yr, above-mentioned specific primer sets, contain the specific primer group
The kit of conjunction is improved in Wheat Germplasm Resources or in the initiative of wheat stripe rust resisting disease material or in wheat marker assisted selection
Application in.
The present invention provides above-mentioned molecular labeling KASP-Yr, above-mentioned specific primer sets, contain the specific primer group
Application of the kit of conjunction in identification stripe rust resisting wheat or the wheat of cultivation stripe rust resisting character.
The present invention provides above-mentioned molecular labeling KASP-Yr, above-mentioned specific primer sets, contain the specific primer group
Application of the kit of conjunction in identification wheat stripe rust resisting ospc gene QYr.sicau-1B.1.
The above-mentioned application of the present invention uses quantitative fluorescent PCR specially using the genomic DNA of Plant samples to be measured as template
Primer carries out fluorescent quantitative PCR, carries out genotyping using amplification, will read primer SEQ ID
The plant for the fluorescence that NO.2 is modified is accredited as the plant containing wheat stripe rust resisting disease QTL QYr.sicau-1B.1.
The present invention provides it is a kind of identify wheat stripe rust resisting disease QTL QYr.sicau-1B.1 molecule labelling method, with to
The DNA of expert evidence as template, with three primer sequences respectively the specific primer as shown in NO.1~3 SEQ ID into
Row PCR amplification simultaneously reads fluorescent value, if there is the fluorescence that SEQ ID NO.2 is modified, can determine whether to there is stripe rust resisting QTL
The wheat of QYr.sicau-1B.1.
Specifically, above-mentioned application, includes the following steps:
1) genomic DNA of plant to be measured is extracted；
2) using the genomic DNA of plant to be measured as template, using the primer of amplifier molecule label KASP-Yr, in instrument
CFX96Real-Time System carries out pcr amplification reaction and reads fluorescent value；
3) pcr amplification product fluorescence is detected, if it is possible to read HEX fluorescence, then plant to be measured is with Rust resistance characteristic of disease
The wheat resource of shape.
The amplification system of above-mentioned PCR amplification are as follows: 5 μ L Master Mix, three primer SEQ ID No:1,2 and 3 according to
The concentration of 10ng/ μ L is separately added into 120 μ L, 120 μ L and 300 μ L and adds after 460 μ L of ddH2O is mixed as mixing
Primer uses, and it is 10 μ L that 1.4 μ L of mix primer, 5ng template DNA, distilled water, which add to total amount, while need to add at least three independence
With distilled water replace DNA profiling blank.
The program of above-mentioned PCR amplification are as follows: 94 DEG C of initial denaturation 15min；94 DEG C of denaturation 20s, 61 DEG C of renaturation/extension 60s, altogether
10 circulations；94 DEG C of denaturation 20s, 55 DEG C of renaturation/extension 60s, totally 26 recycle；Fluorescence readings is carried out after the completion.
The invention discloses be located on wheat 1B chromosome with the wheat stripe rust resisting chain molecular labeling KASP-Yr of disease,
The molecular labeling is the flanking marker of stripe rust resisting QTL QYr.sicau-1B.1 on wheat 1B chromosome long arm, and chain degree is high.
The label can be used to detect the stripe rust resisting QTL on wheat 1B chromosome, and quickly screening has the plant in the site, Jin Erfang
Just the marker assisted selection of highly resistance wheat is carried out.Stripe rust resisting on molecular labeling KASP-Yr provided by the invention and wheat 1B
QTL QYr.sicau-1B.1 close linkage can be used to position this sick character of wheat stripe rust resisting, thus in breeding
Susceptible plant is eliminated in journey, improves breeding work efficiency, and the research for wheat stripe rust resisting ospc gene provides basis.
Present invention firstly discloses the stripe rust resisting QTL QYr.sicau-1B.1 for coming from wheat ' 20828 ', are located at wheat
On 1B chromosome long arm, wheat stripe rust resisting is dramatically increased.The QTL has in disease-resistant wheat (regulation strip rust resistemce) breeding
Higher utility value.
Present invention firstly discloses the stripe rust resisting QTL that wheat ' 20828 ' is accurately detected based on quantitative fluorescent PCR platform
The molecular labeling KASP-Yr of QYr.sicau-1B.1, and be codominant marker, detection precise and high efficiency, amplification facilitate stabilization.
Molecular labeling KASP-Yr disclosed by the invention and stripe rust resisting QTL QYr.sicau-1B.1 are extremely significant related, are in
Marker characteristic is now isolated, the accuracy for molecular marker assisted selection is high, improves the wheat stripe rust resisting for adapting to varying environment
The selection determination rates of sick kind, and success rate is high.
Detailed description of the invention
Fig. 1 is that wheat stripe rust resisting QTL QYr.sicau-1B.1 determines on 1B chromosome in the embodiment of the present invention 1
Position.
Fig. 2 is the recombinant inbred lines strain plant molecular labeling KASP- of ' 20828 ' × ' CN16 ' in the embodiment of the present invention 1
The fluorescence readings result of Yr detection；Wherein, HEX (blue, ' 20828 ') fluorescence is the strain of stripe rust resisting, FAM (it is orange,
' CN16 ') fluorescence be susceptible strain；Green fluorescence is heterozygosis strain；Black fluorescent is blank control.
Fig. 3 is that the recombinant inbred lines strain of ' 20828 ' × common wheat strain ' SY95-71 ' in the embodiment of the present invention 2 is planted
Strain molecule marks the fluorescence readings result of KASP-Yr detection；Wherein, HEX (blue, ' 20828 ') fluorescence is the strain of stripe rust resisting
System, FAM (orange, ' SY95-71 ') fluorescence are susceptible strain；Green fluorescence is heterozygosis strain；Black fluorescent is blank control.
Specific embodiment
Following embodiment further illustrates the contents of the present invention, but should not be construed as limiting the invention.Without departing substantially from
In the case where spirit of that invention and essence, to modifications or substitutions made by the method for the present invention, step or condition, the present invention is belonged to
Range.
Wheat Germplasm Resources used in the embodiment of the present invention are all from Triticeae Research Institute, Sichuan Agricultural University germ plasm resource
Library.
Unless otherwise specified, the conventional means that technological means used in embodiment is well known to those skilled in the art.
The acquisition of embodiment 1 wheat stripe rust resisting ospc gene QYr.sicau-1B-1 and its molecular labeling KASP-Yr
(1) using wheat line ' 20828 ' to be maternal, with wheat breed ' CN16 ' for paternal hybrid, hybrid F is obtained1, F1
F is obtained for individual plant selfing2, in F2Using Dan Suichuan method, until F8In generation, obtains the recombinant inbred lines for being containing 199, constitutes
Genetic mapping group.
(2) recombinant inbred lines stripe rust phenotypic evaluation is after the onset of inducing materials SY95-71 is abundant, field record
The disease resistance of material to be tested is reacted, and response type determines each strain in group according to 1-9 grades of classification standards, determines knot
Fruit is denoted as the morphological markers data of Stripe Rust Resistance Gene QYr.sicau-1B.1.The position navigated on linkage map in this way is exactly
QYr.sicau-1B.1 gene position.
(3) 55K SNP chip is analyzed
A) DNA is extracted: extracting parent ' 20828 ', ' CN16 ' and recombinant inbred lines plant DNA with CTAB method.
B) quality testing is carried out using DNA of the ultramicrospectrophotometer to extraction, sample presentation to company carries out base after qualification
Because of type analysis, the genotyping of parents and mapping population is by Beijing Bo Aojing allusion quotation Bioisystech Co., Ltd in our current research
The 55K SNP chip of (http://www.capitalbiotech.com) and Jia Jizeng seminar cooperative development is completed, the chip
It is commercially available.
C) building of linkage map: according to 55K SNP chip data, genetic map is constructed using JoinMap4.0.In conjunction with
The stripe rust phenotypic data of group, with the complete Interval Mapping (Inclusive in QTL IciMapping 4.0
Composite Interval Mapping-ADD, ICIM-ADD), it is arranged under conditions of threshold values LOD >=2.5, uses 2016-
2018 3 times totally 4 ecosites and BLUP (optimal linear unbiased prediction, the best linear of 4 ecosite stripe rust
Unbiased prediction) value detects QTL, wheat stripe rust resisting disease QTL QYr.sicau-1B.1 is oriented, and calculate
Genetic distance between the position and molecular labeling of QYr.sicau-1B.1.
D) it the acquisition of the densification of genetic map and compact linkage molecule label: for densification genetic map and obtains and anti-item
The molecular labeling of rust QTL QYr.sicau-1B.1 close linkage, using 55K SNP chip data positioning result to flank mark
It remembers row physical positioning into and screens the gene being located in section, these genes are cloned in parent ' 20828 ' and ' CN16 ',
To obtaining polymorphic site and carrying out the exploitation of molecular labeling, DNAMAN design KASP primer is utilized (to design 30,10 pairs altogether
KASP primer) (table 1), finally obtain label KASP-Yr and stripe rust resisting QTL QYr.sicau-1B.1 close linkage, the SNP
Molecular labeling be averaged LOD value be 21.39, explain about 25.52% phenotypic variation.
E) it is analyzed.1 molecular labeling and stripe rust resisting QTL have been finally obtained in 10 pairs of KASP primers of design
QYr.sicau-1B.1 close linkage.The result is shown in Figure 1,2.
Application of the 2 molecular labeling KASP-Yr of embodiment on selection control stripe rust resisting QTL QYr.sicau-1B.1
It (1) is female parent, susceptible common wheat strain ' SY95- using the common wheat strain ' 20828 ' of stripe rust resisting
71 ' construct recombinant inbred lines for male parent, and 163 strains are randomly choosed in offspring's strain.
(2) KASP-Yr label detection is carried out to 163 strains obtained, method particularly includes: extract 163 strains
DNA；As template, with the specific primer of molecular labeling KASP-Yr to carrying out PCR amplification for primer and carry out fluorescence
Readings, the label KASP-Yr primer are as follows:
Primer on FAM label: (underscore part is FAM sequence label)
5’-GAAGGTGACCAAGTTCATGCTCTTGTGTGTGACTGTACCATA -3’(SEQ ID No.1)
Primer on HEX label: (glissade part is HEX sequence label)
5’-GAAGGTCGGAGTCAACGGATTCTTGTGTGTGACTGTACCAT G-3’(SEQ ID No.2)
General reverse primer:
5’-CTGATACAGATAGGTAGCAT-3’(SEQ ID No.3)
The amplification system of above-mentioned PCR amplification are as follows: 5 μ L Master Mix, three primer SEQ ID No:1,2 and 3 according to
The concentration of 10ng/ μ L is separately added into 120 μ L, 120 μ L and 300 μ L and adds after 460 μ L of ddH2O is mixed as mixing
Primer uses, and it is 10 μ L that 1.4 μ L of mix primer, 5ng template DNA, distilled water, which add to total amount, while need to add at least three independence
With distilled water replace DNA profiling blank.
The program of above-mentioned PCR amplification are as follows: 94 DEG C of initial denaturation 15min；94 DEG C of denaturation 20s, 61 DEG C of renaturation/extension 60s, altogether
10 circulations；94 DEG C of denaturation 20s, 55 DEG C of renaturation/extension 60s, totally 26 recycle；Fluorescence readings is carried out after the completion.
Fluorescence readings result (see Fig. 3) will test the plant genotype with ' 20828 ' consistent HEX (blue) fluorescence
It is denoted as A, is stripe rust resisting strain, B is denoted as with ' SY95-71 ' plant genotype for equally showing as FAM (orange) fluorescence, for sense
Sick type strain.The BLUP value of each strain genotype and 4 ecosite stripe rust is as shown in table 2.Label KASP-Yr in contain
Having the identical plant mean disease index of ' 20828 ' types of stripe rust resisting QTL QYr.sicau-1B.1 is 37.56, extremely significant
Lower than the plant disease index (average 65.35) with ' SY95-71 ' type.Actual result is consistent with expected results, illustrates this hair
Bright stripe rust resisting QTL QYr.sicau-1B.1 plays the role of significantly increasing stripe rust resisting really；Molecule mark of the invention simultaneously
Remember that KASP-Yr can identify stripe rust resisting QTL QYr.sicau-1B.1 with tracking.
Table 2 ' 20828 ' × ' SY95-71 ' recombinant inbred lines KASP-Yr genotype result corresponding with phenotype
Although having used general explanation, specific embodiment and test above, the present invention is described in detail,
But on the basis of the present invention, it can be made it is some modify or improve, this is apparent to those skilled in the art
's.Therefore, these modifications or improvements without departing from theon the basis of the spirit of the present invention, belong to claimed
Range.
Sequence table
<110>Sichuan Agricultural University
<120>with wheat stripe rust resisting ospc gene QYr.sicau-1B-1 chain SNP marker and application
<130> KHP181115919.6
<160> 31
<170> SIPOSequenceListing 1.0
<210> 1
<211> 21
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 1
cttgtgtgtg actgtaccat a 21
<210> 2
<211> 21
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 2
cttgtgtgtg actgtaccat g 21
<210> 3
<211> 20
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 3
ctgatacaga taggtagcat 20
<210> 4
<211> 21
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 4
ccgagcagat ggttaaggct a 21
<210> 5
<211> 21
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 5
ccgagcagat ggttaaggct c 21
<210> 6
<211> 20
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 6
gcgagagttc tcggcgttga 20
<210> 7
<211> 21
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 7
gcatgcaacc tagatgttag g 21
<210> 8
<211> 21
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 8
gcatgcaacc tagatgttag t 21
<210> 9
<211> 20
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 9
agaaaaataa tgcttcctcg 20
<210> 10
<211> 21
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 10
tgacgaagaa ctagtacaat g 21
<210> 11
<211> 21
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 11
tgacgaagaa ctagtacaat t 21
<210> 12
<211> 21
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 12
gaacacctct cctccggagg c 21
<210> 13
<211> 21
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 13
tacctgtcgg tatgtctact c 21
<210> 14
<211> 21
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 14
tacctgtcgg tatgtctact t 21
<210> 15
<211> 20
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 15
ttcctctggc ttgatattca 20
<210> 16
<211> 21
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 16
cagaacaatt atcatgtagc g 21
<210> 17
<211> 21
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 17
cagaacaatt atcatgtagc t 21
<210> 18
<211> 20
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 18
tgcactggtt ctccagttat 20
<210> 19
<211> 21
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 19
gaggttaact ccttcctctt c 21
<210> 20
<211> 21
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 20
gaggttaact ccttcctctt t 21
<210> 21
<211> 20
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 21
ttcttatgtg aatctgaagg 20
<210> 22
<211> 21
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 22
gtcctgcgtc ggatccgtgg c 21
<210> 23
<211> 21
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 23
gtcctgcgtc ggatccgtgg t 21
<210> 24
<211> 20
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 24
tggattacaa gctaccatat 20
<210> 25
<211> 19
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 25
gtattatctt gtaatccta 19
<210> 26
<211> 19
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 26
gtattatctt gtaatcctg 19
<210> 27
<211> 20
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 27
aaggtccata tataatgata 20
<210> 28
<211> 20
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 28
tctctcatgc attgtatgca 20
<210> 29
<211> 20
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 29
tctctcatgc attgtatgcg 20
<210> 30
<211> 20
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 30
tcatcgagac gaccaaccaa 20
<210> 31
<211> 71
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 31
tatgctagtg ctatgcttgt gtgtgactgt accatacttt cttatcatgc tacctatctg 60
tatcagttat g 71
Claims (10)
1. with wheat stripe rust resisting ospc gene QYr.sicau-1B.1 chain SNP marker, which is characterized in that the molecule mark
It is denoted as KASP-Yr, the molecular labeling is long in wheat 1B chromosome with wheat stripe rust resisting disease QTL QYr.sicau-1B.1 common location
Genetic distance on arm, and between QYr.sicau-1B.1 is less than 1.3cM, and the polymorphism of the SNP marker is A/G.
2. SNP marker as described in claim 1, which is characterized in that it can be obtained by following 3 primer amplifications: institute
Stating 3 primers includes 2 upstream primers and 1 general reverse primer, and 2 upstream primers 5 ' end is modified different glimmering respectively
Light group；The nucleotide sequence of 3 primers is respectively as shown in SEQ ID NO.1,2 and 3.
3. application of the wheat stripe rust resisting disease QTL QYr.sicau-1B.1 in regulation wheat stripe rust resisting characteristic of disease shape, the wheat
Stripe rust resisting QTL QYr.sicau-1B.1, from female parent ' 20828 ', the QTL be located at chromosome of wheat 1B it is long-armed on,
The physical location of RefSeqv1.0 genome version is 474Mb.
4. the specific primer group for detecting the chain SNP marker of wheat stripe rust resisting disease QTL QYr.sicau-1B.1
It closes, which is characterized in that contain nucleotide sequence primer as shown in NO.1~3 SEQ ID.
5. the kit containing specific primer sets described in claim 4.
6. any SNP marker of claim 1-2 or specific primer sets as claimed in claim 4 or right are wanted
Application of the kit described in asking 5 in identification wheat stripe rust resisting ospc gene QYr.sicau-1B.1.
7. any SNP marker of claim 1-2 or specific primer sets as claimed in claim 4 or right are wanted
Application of the kit described in asking 5 in identification stripe rust resisting wheat.
8. any SNP marker of claim 1-2 or specific primer sets as claimed in claim 4 or right are wanted
Kit described in asking 5 is improved in Wheat Germplasm Resources or in the initiative of wheat stripe rust resisting disease material or in wheat marker assisted selection
In application.
9. such as application as claimed in claim 6 to 8, which is characterized in that using the genomic DNA of Plant samples to be measured as mould
Plate carries out fluorescent quantitative PCR with fluorescence quantification PCR primer, carries out genotyping using amplification, will read
The plant for the fluorescence modified to primer SEQ ID NO.2 is accredited as containing wheat stripe rust resisting disease QTL QYr.sicau-1B.1's
Plant.
10. application as claimed in claim 9, which is characterized in that the amplification reaction system of the quantitative fluorescent PCR: 5 μ L
Master Mix, 3 primer SEQ ID NO.1,2 and 3 are separately added into 120 μ L, 120 μ L and 300 μ according to the concentration of 10ng/ μ L
L and adding is used after 460 μ L of ddH2O is mixed as mix primer, 1.4 μ L of mix primer, 5ng template DNA, distilled water
Adding to total amount is 10 μ L, while need to add the independent blank that DNA profiling is replaced with distilled water of at least three；
Quantitative fluorescent PCR program: 94 DEG C of initial denaturation 15min；94 DEG C of denaturation 20s, 61 DEG C of renaturation/extension 60s, totally 10 recycle；
94 DEG C of denaturation 20s, 55 DEG C of renaturation/extension 60s, totally 26 recycle；
The result of acquisition carries out Genotyping；Detect pcr amplification product fluorescence, if it is possible to read primer SEQ ID NO.2 institute
The fluorescence of modification, then plant to be measured is the wheat resource with stripe rust resisting character.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910134070.9A CN109706263B (en) | 2019-02-22 | 2019-02-22 | SNP molecular marker linked with wheat stripe rust resistance gene QYr. sicau-1B-1 and application |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910134070.9A CN109706263B (en) | 2019-02-22 | 2019-02-22 | SNP molecular marker linked with wheat stripe rust resistance gene QYr. sicau-1B-1 and application |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN109706263A true CN109706263A (en) | 2019-05-03 |
| CN109706263B CN109706263B (en) | 2022-04-05 |
Family
ID=66263869
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201910134070.9A Active CN109706263B (en) | 2019-02-22 | 2019-02-22 | SNP molecular marker linked with wheat stripe rust resistance gene QYr. sicau-1B-1 and application |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN109706263B (en) |
Cited By (20)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110499387A (en) * | 2019-09-27 | 2019-11-26 | 四川农业大学 | A kind of molecular labeling and its application that the long QTL of wheat flag leaf is chain |
| CN111073993A (en) * | 2020-01-08 | 2020-04-28 | 中国农业大学 | SNP site of wheat leaf rust resistance gene Lr13 and application thereof |
| CN111471790A (en) * | 2020-04-26 | 2020-07-31 | 四川农业大学 | Molecular marker closely linked with wheat grain filling rate QT L QGfr. sicau-7D.1 and application thereof |
| CN111575400A (en) * | 2020-06-28 | 2020-08-25 | 湖北省农业科学院粮食作物研究所 | Wheat stripe rust resistant QTL molecular marker IWB12253 and application thereof |
| CN111607664A (en) * | 2020-07-13 | 2020-09-01 | 湖北省农业科学院粮食作物研究所 | Application of SNP molecular marker on 1DS chromosome related to wheat stripe rust |
| CN111635957A (en) * | 2020-07-20 | 2020-09-08 | 湖北省农业科学院粮食作物研究所 | Molecular marker for detecting wheat stripe rust resistance QTL and application of molecular marker in disease-resistant breeding |
| CN111778346A (en) * | 2020-07-06 | 2020-10-16 | 湖北省农业科学院粮食作物研究所 | Molecular marker for detecting stripe rust resistant QTL QYr. hbaas-4BL.1 and using method thereof |
| CN112430606A (en) * | 2020-12-11 | 2021-03-02 | 四川农业大学 | Wheat stripe rust resistant gene yrZ15-1949 and molecular marker and application thereof |
| CN113046467A (en) * | 2020-12-23 | 2021-06-29 | 山东农业大学 | SNP loci significantly associated with wheat stripe rust resistance and application thereof in genetic breeding |
| CN113388697A (en) * | 2021-08-05 | 2021-09-14 | 西北农林科技大学 | KASP-SNP molecular probe and detection method and application thereof |
| CN113637789A (en) * | 2021-08-13 | 2021-11-12 | 四川农业大学 | Wheat stripe rust resistant gene YRTD121 linked KASP molecular marker, primer, kit and application |
| CN113755637A (en) * | 2021-10-18 | 2021-12-07 | 四川农业大学 | Wheat speckle leaf gene Lm5 coseparated KASP molecular marker and application thereof |
| CN113897457A (en) * | 2021-11-29 | 2022-01-07 | 四川农业大学 | KASP molecular marker linked with wheat stripe rust resistant QTL and application thereof |
| CN114410652A (en) * | 2022-01-21 | 2022-04-29 | 西南科技大学 | Stripe rust resistant gene QYRsv and swust-1BL tightly linked molecular marker in adult plant stage and application thereof |
| CN114836556A (en) * | 2022-03-24 | 2022-08-02 | 四川农业大学 | Molecular marker closely linked with wheat stripe rust resistance QTL QYR |
| CN115896339A (en) * | 2023-02-14 | 2023-04-04 | 四川农业大学 | Specific SNP molecular marker related to wheat stripe rust resistance gene Yr81 and application thereof |
| CN115927726A (en) * | 2022-11-14 | 2023-04-07 | 四川农业大学 | KASP specific primer of SNP molecular marker linked with wheat stripe rust resistance gene and application |
| CN116024379A (en) * | 2023-02-14 | 2023-04-28 | 西北农林科技大学 | Wheat stripe rust resistance gene YrAK58 functional molecular marker and application thereof |
| CN116121444A (en) * | 2023-03-03 | 2023-05-16 | 西北农林科技大学 | Functional KASP molecular marker of wheat stripe rust resistance gene YrNP63 and application thereof |
| CN118421830A (en) * | 2024-07-04 | 2024-08-02 | 四川省农业科学院作物研究所(四川省种质资源中心) | KASP molecular marker linked with Yizhang red shell wheat stripe rust resistance gene QYr.YZHK-2BL, primer combination and application |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20140359898A1 (en) * | 2013-06-03 | 2014-12-04 | The Board Of Regents For Oklahoma State University | Transgenic cereal plants with increased resistance to rust diseases |
| CN105256044A (en) * | 2015-11-03 | 2016-01-20 | 中国农业科学院作物科学研究所 | Wheat molecular barcode based on single nucleotide polymorphism |
| CN109321671A (en) * | 2018-10-10 | 2019-02-12 | 四川农业大学 | Wheat stripe rust resisting ospc gene QYr.sicau-1B-1 SSR molecular marker and application |
-
2019
- 2019-02-22 CN CN201910134070.9A patent/CN109706263B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20140359898A1 (en) * | 2013-06-03 | 2014-12-04 | The Board Of Regents For Oklahoma State University | Transgenic cereal plants with increased resistance to rust diseases |
| CN105256044A (en) * | 2015-11-03 | 2016-01-20 | 中国农业科学院作物科学研究所 | Wheat molecular barcode based on single nucleotide polymorphism |
| CN109321671A (en) * | 2018-10-10 | 2019-02-12 | 四川农业大学 | Wheat stripe rust resisting ospc gene QYr.sicau-1B-1 SSR molecular marker and application |
Non-Patent Citations (3)
| Title |
|---|
| JIAN MA等: "Identification and validation of a novel major QTL for all-stage stripe rust resistance on 1BL in the winter wheat line 20828", 《THEORETICAL AND APPLIED GENETICS》 * |
| 伍玲: "小麦条锈病成株抗性QTL定位和分子标记检测", 《中国博士学位论文全文数据库农业科技辑》 * |
| 黄亮等: "中国79个小麦品种（系）抗条锈病评价及基因分子检测", 《中国农业科学》 * |
Cited By (29)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110499387A (en) * | 2019-09-27 | 2019-11-26 | 四川农业大学 | A kind of molecular labeling and its application that the long QTL of wheat flag leaf is chain |
| CN110499387B (en) * | 2019-09-27 | 2021-12-03 | 四川农业大学 | Wheat flag leaf length QTL linked molecular marker and application thereof |
| CN111073993A (en) * | 2020-01-08 | 2020-04-28 | 中国农业大学 | SNP site of wheat leaf rust resistance gene Lr13 and application thereof |
| CN111073993B (en) * | 2020-01-08 | 2022-01-18 | 中国农业大学 | SNP site of wheat leaf rust resistance gene Lr13 and application thereof |
| CN111471790A (en) * | 2020-04-26 | 2020-07-31 | 四川农业大学 | Molecular marker closely linked with wheat grain filling rate QT L QGfr. sicau-7D.1 and application thereof |
| CN111575400A (en) * | 2020-06-28 | 2020-08-25 | 湖北省农业科学院粮食作物研究所 | Wheat stripe rust resistant QTL molecular marker IWB12253 and application thereof |
| CN111575400B (en) * | 2020-06-28 | 2023-04-25 | 湖北省农业科学院粮食作物研究所 | Wheat stripe rust resistance QTL molecular marker IWB12253 and application thereof |
| CN111778346A (en) * | 2020-07-06 | 2020-10-16 | 湖北省农业科学院粮食作物研究所 | Molecular marker for detecting stripe rust resistant QTL QYr. hbaas-4BL.1 and using method thereof |
| CN111778346B (en) * | 2020-07-06 | 2023-09-05 | 湖北省农业科学院粮食作物研究所 | Molecular marker for detecting stripe rust resistance QTL QYr.hbas-4BL.1 and using method |
| CN111607664B (en) * | 2020-07-13 | 2023-08-08 | 湖北省农业科学院粮食作物研究所 | Application of SNP molecular marker on 1DS chromosome related to wheat stripe rust |
| CN111607664A (en) * | 2020-07-13 | 2020-09-01 | 湖北省农业科学院粮食作物研究所 | Application of SNP molecular marker on 1DS chromosome related to wheat stripe rust |
| CN111635957B (en) * | 2020-07-20 | 2023-06-30 | 湖北省农业科学院粮食作物研究所 | Molecular marker for detecting wheat stripe rust resistance QTL and application of molecular marker in disease resistance breeding |
| CN111635957A (en) * | 2020-07-20 | 2020-09-08 | 湖北省农业科学院粮食作物研究所 | Molecular marker for detecting wheat stripe rust resistance QTL and application of molecular marker in disease-resistant breeding |
| CN112430606A (en) * | 2020-12-11 | 2021-03-02 | 四川农业大学 | Wheat stripe rust resistant gene yrZ15-1949 and molecular marker and application thereof |
| CN113046467A (en) * | 2020-12-23 | 2021-06-29 | 山东农业大学 | SNP loci significantly associated with wheat stripe rust resistance and application thereof in genetic breeding |
| CN113388697A (en) * | 2021-08-05 | 2021-09-14 | 西北农林科技大学 | KASP-SNP molecular probe and detection method and application thereof |
| CN113637789A (en) * | 2021-08-13 | 2021-11-12 | 四川农业大学 | Wheat stripe rust resistant gene YRTD121 linked KASP molecular marker, primer, kit and application |
| CN113755637A (en) * | 2021-10-18 | 2021-12-07 | 四川农业大学 | Wheat speckle leaf gene Lm5 coseparated KASP molecular marker and application thereof |
| CN113897457A (en) * | 2021-11-29 | 2022-01-07 | 四川农业大学 | KASP molecular marker linked with wheat stripe rust resistant QTL and application thereof |
| CN114410652A (en) * | 2022-01-21 | 2022-04-29 | 西南科技大学 | Stripe rust resistant gene QYRsv and swust-1BL tightly linked molecular marker in adult plant stage and application thereof |
| CN114836556A (en) * | 2022-03-24 | 2022-08-02 | 四川农业大学 | Molecular marker closely linked with wheat stripe rust resistance QTL QYR |
| CN114836556B (en) * | 2022-03-24 | 2023-08-29 | 四川农业大学 | Molecular marker closely linked with wheat stripe rust resistance QTL QYr.sicau-6B and application |
| CN115927726A (en) * | 2022-11-14 | 2023-04-07 | 四川农业大学 | KASP specific primer of SNP molecular marker linked with wheat stripe rust resistance gene and application |
| CN115896339A (en) * | 2023-02-14 | 2023-04-04 | 四川农业大学 | Specific SNP molecular marker related to wheat stripe rust resistance gene Yr81 and application thereof |
| CN116024379A (en) * | 2023-02-14 | 2023-04-28 | 西北农林科技大学 | Wheat stripe rust resistance gene YrAK58 functional molecular marker and application thereof |
| CN116024379B (en) * | 2023-02-14 | 2024-04-12 | 西北农林科技大学 | Wheat stripe rust resistance gene YrAK58 functional molecular marker and application thereof |
| CN116121444A (en) * | 2023-03-03 | 2023-05-16 | 西北农林科技大学 | Functional KASP molecular marker of wheat stripe rust resistance gene YrNP63 and application thereof |
| CN118421830A (en) * | 2024-07-04 | 2024-08-02 | 四川省农业科学院作物研究所(四川省种质资源中心) | KASP molecular marker linked with Yizhang red shell wheat stripe rust resistance gene QYr.YZHK-2BL, primer combination and application |
| CN118421830B (en) * | 2024-07-04 | 2024-09-06 | 四川省农业科学院作物研究所(四川省种质资源中心) | KASP molecular marker linked with Yizhang red shell wheat stripe rust resistance gene QYr.YZHK-2BL, primer combination and application |
Also Published As
| Publication number | Publication date |
|---|---|
| CN109706263B (en) | 2022-04-05 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN109706263A (en) | Chain SNP marker and application with wheat stripe rust resisting ospc gene QYr.sicau-1B-1 | |
| CN109825621A (en) | Wheat spikelet number QTL chain SNP marker and its application | |
| CN106868131B (en) | SNP molecular marker of upland cotton No. 6 chromosome related to fiber strength | |
| CN110295251A (en) | Chain SNP marker and its application with wheat available tillering QTL | |
| CN110499387B (en) | Wheat flag leaf length QTL linked molecular marker and application thereof | |
| CN112593007B (en) | SNP molecular marker linked with wheat grain length QTL and application thereof | |
| CN112481275A (en) | Wheat stripe rust resistant gene yrZ15-1370 and molecular marker and application thereof | |
| CN114774570B (en) | Molecular marker closely linked with wheat stem rot resistance QTL and application | |
| CN109929945B (en) | Molecular marker BrSF2604 primer of main effect QTL sites in flowering phase and mature phase of brassica napus and application thereof | |
| CN115786565B (en) | SNP molecular marker linked with wheat spike number QTL and application thereof | |
| CN108374054A (en) | Suitable for one group of rice SSR molecular marker of capillary electrophoresis detection technology and its application | |
| CN108179220B (en) | KASP marker tightly linked with wheat dwarf gene Rht12 and application thereof | |
| CN111471790B (en) | Molecular marker closely linked with wheat grain filling rate QTL QGfr. sicau-7D.1 and application thereof | |
| CN112779350A (en) | Molecular marker closely linked with wheat spikelet grain number QTLQGns | |
| CN103146831B (en) | Method for identifying rice and dry rice | |
| CN105296472B (en) | The molecular labeling and its screening technique in Sand Pear ' faint scent ' pericarp brown character gene site entirely | |
| CN111647677B (en) | Molecular marker closely linked with wheat grain filling rate QTL QGfr. sicau-6D and application | |
| CN107955838A (en) | The exploitation and application of thcrmo-scnsitivc genie male stcrility gene TMS5 Functional markers | |
| CN110283929A (en) | The relevant SNP marker 5-160 of capsicum epidemic disease resistant gene and its specific primer and application | |
| CN113215297B (en) | Molecular marker ID0159 closely linked with major QTL site of sesame oil content and application thereof | |
| CN108624710A (en) | A kind of and the relevant SSR marker of cucumber fruit length character and its application | |
| CN105524994B (en) | The molecular labeling HRM7 of barley grain length gene Lkl2 a kind of and its application | |
| CN111100946B (en) | Molecular marker primer of rape grain weight character major gene locus and application | |
| CN108060247B (en) | Haplotype related to upland cotton No. 8 chromosome fiber strength | |
| CN111485032A (en) | Method for identifying cucumber female line and SNP primer combination used by same |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |