CN107794308A - Identify special SNP and its application of wheat seed character - Google Patents
Identify special SNP and its application of wheat seed character Download PDFInfo
- Publication number
- CN107794308A CN107794308A CN201610757883.XA CN201610757883A CN107794308A CN 107794308 A CN107794308 A CN 107794308A CN 201610757883 A CN201610757883 A CN 201610757883A CN 107794308 A CN107794308 A CN 107794308A
- Authority
- CN
- China
- Prior art keywords
- wheat
- genotype
- sequence
- kernel
- mass
- 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
Classifications
-
- 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
-
- 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
-
- 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
Landscapes
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Analytical Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Organic Chemistry (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Health & Medical Sciences (AREA)
- Biotechnology (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Immunology (AREA)
- Mycology (AREA)
- Microbiology (AREA)
- Molecular Biology (AREA)
- Botany (AREA)
- Biophysics (AREA)
- Physics & Mathematics (AREA)
- Biochemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The invention discloses a kind of special SNP for identifying wheat seed character and its application.The invention provides a kind of method for identifying or assisting in discriminating wheat seed character, comprise the following steps：It is AA genotype, AT genotype or TT genotype to detect the genotype based on 2144SNP sites in the genomic DNA of wheat to be measured；The grain characters of AA genotype wheats are better than TT genotype wheats；The grain characters are excellent to be presented as that mass of 1000 kernel is high and/or grain length is grown.The present invention develops the SNP site related to wheat seed character, and the primer sets based on KASP technologies are devised on the basis of the SNP site.Wheat seed character is identified using primer sets provided by the invention, it is quick and easy, accurate to have the advantages that.The present invention is for wheat of the seed selection with different grain characters with fabulous application prospect.
Description
Technical field
The invention belongs to biological technical field, and in particular to a kind of special SNP for identifying wheat seed character and its application.
Background technology
Wheat is the cereal crops that cultivated area is maximum in the world, in China, the cultivated area of wheat be only second to corn and
Rice, account for 21% or so of total grain output value.Wheat yield is to directly affect our people's living standard height and national food
An important factor for whether safe.Improve wheat yield, make its stable high yield be that China wheat breeding men chase after for a long time all the time
The main target asked.However, China's Wheat Production is in slow developing stage at present, yield of wheat increases less than 0.8% every year.
With the growth of population, urbanization, desertification and salinization of soil, foodstuff planting area will be reduced increasingly, this and grain
The contradiction that demand constantly rigidly increases is more and more prominent and serious.Therefore, wheat yield phase is cloned using Protocols in Molecular Biology
Functional gene is closed, and with this development functionality molecular labeling, important references gene money is provided for wheat molecular marker assistant breeding
Source, there is important scientific meaning and practical application valency to accelerating China's wheat breeding process, improving China's wheat yield aspect
Value.
Grain is one of three elements of yield again, determines the key factor of grain weight and includes the filling rate of grain type and seed.
In production and breeding practice, mass of 1000 kernel is often used as to weigh the index of seed size, mainly by grain type characteristic index (such as grain
Long, grain is wide and the influent factors such as grain is thick) form and with yield significant positive correlation.
The content of the invention
It is an object of the invention to provide a kind of special SNP for identifying wheat seed character and its application.
The invention provides a kind of method for identifying or assisting in discriminating wheat seed character, comprise the following steps：Detection is treated
It is AA genotype, AT genotype or TT genotype to survey the genotype based on 2144 SNP sites in the genomic DNA of wheat；AA
The grain characters of genotype wheat are better than TT genotype wheats；The grain characters are excellent to be presented as that mass of 1000 kernel is high and/or grain length is grown.
Present invention also offers a kind of method identified or aid in identification wheat seed mass of 1000 kernel, comprise the following steps：Inspection
It is AA genotype, AT genotype or TT genes to survey the genotype based on 2144 SNP sites in the genomic DNA of wheat to be measured
Type；It is high mass of 1000 kernel wheat if AA genotype, wheat candidate to be measured；It is low if TT genotype, wheat candidate to be measured
Mass of 1000 kernel wheat；The high mass of 1000 kernel wheat is seed mass of 1000 kernel >=35g wheat；The low mass of 1000 kernel wheat is seed thousand
Weight ＜ 35g wheat.
Present invention also offers a kind of method identified or aid in identification wheat seed grain length, comprise the following steps：Detection
Genotype based on 2144 SNP sites in the genomic DNA of wheat to be measured is AA genotype, AT genotype or TT genotype；
It is long grain length wheat if AA genotype, wheat candidate to be measured；It is that short grain length is small if TT genotype, wheat candidate to be measured
Wheat；The long grain length wheat is seed grain length >=0.65mm wheat；The short grain length wheat is seed grain length ＜ 0.65mm's
Wheat.
In any of the above methods described, " genotype based on 2144 SNP sites in the genomic DNA of wheat to be measured is detected
For AA genotype, AT genotype or TT genotype " method comprise the following steps：Using the genomic DNA of wheat to be measured as mould
Plate, performing PCR amplification is entered using special primer group, genotype results are obtained by detecting pcr amplification product.
The special primer group is primer sets I or primer sets II；
The primer sets I are made up of 2144F1,2144F2 and 2144C；
The primer 2 144F1 is following (b1) or (b2)：
(b1) single strand dna shown in the sequence 5 of sequence table；
(b2) sequence 5 is had by the substitution of one or several nucleotides and/or missing and/or addition and with sequence 5
The DNA molecular of identical function；
The 2144F2 is following (b3) or (b4)：
(b3) single strand dna shown in the sequence 6 of sequence table；
(b4) sequence 6 is had by the substitution of one or several nucleotides and/or missing and/or addition and with sequence 6
The DNA molecular of identical function；
The 2144C is following (b5) or (b6)：
(b5) single strand dna shown in the sequence 7 of sequence table；
(b6) sequence 7 is had by the substitution of one or several nucleotides and/or missing and/or addition and with sequence 7
The DNA molecular of identical function.
The primer sets II are made up of TaTPP-F1 and TaTPP-R1；
The TaTPP-F1 is following (c1) or (c2)：
(c1) single strand dna shown in the sequence 1 of sequence table；
(c2) sequence 1 is had by the substitution of one or several nucleotides and/or missing and/or addition and with sequence 1
The DNA molecular of identical function；
The TaTPP-R1 is following (c3) or (c4)：
(c3) single strand dna shown in the sequence 2 of sequence table；
(c4) sequence 2 is had by the substitution of one or several nucleotides and/or missing and/or addition and with sequence 2
The DNA molecular of identical function.
The primer sets I are the primer sets based on KASP technologies.
The primer sets II are the primer sets based on regular-PCR technology.
When using the primer sets I, in the reaction system of PCR amplifications, 2144F1,2144F2 and 2144C mol ratio
For 12：12：30.
When using the primer sets I, in the reaction system of PCR amplifications, 2144F1 concentration is 12 μM, 2144F2 it is dense
The concentration spent for 12 μM, 2144C is 30 μM.
When using the primer sets I, in the reaction system of PCR amplifications, the concentration of magnesium ion is 0.2mM.
When using the primer sets I, the reaction system composition of PCR amplifications is concretely：(DNA content is about by the μ L of template 2.2
For 100ng), MgCl2μ L of the aqueous solution 0.04, μ L of 2 × Master Mix 2.5, (mix primer solution contains mix primer solution
2144F1,2144F2 and 2144C) 0.056 μ L, use ddH2O complements to 5 μ L.
2 × Master Mix full name is " KASP 2 × Master of V4.0 Mix 96/384 (Low Rox) ", Beijing
LGC companies, catalog number KBS-1016-017.
When using the primer sets I, the response procedures of PCR amplifications are concretely：94℃15min；It is 95 DEG C of 20s, a certain
Temperature 20s (temperature of circulation is 65 DEG C for the first time, and each previous circulation of recycle ratio lowers 1 DEG C), 10 circulations；95℃20s、57
DEG C 20s, 30 circulations.
When using the primer sets I, PCR amplifications are run on the instruments of QuantStudio 7 of ABI companies production, from
Dynamic output genotypic results.It is AA genotype for examination wheat if the result of Genotyping is Alle 2 (blue round dot)；Such as
The result of fruit gene parting is Alle 1 (red spots), is TT genotype for examination wheat；If genotypic results are Alle
1/Alle 2, it is AT genotype for examination wheat.
The present invention also protect for detect wheat genomic DNA in genotype based on 2144 SNP sites material
Using for following (d1) or (d2) or (d3) or (d4)：
(d1) identify or assisting in and differentiate wheat seed character；The grain characters are mass of 1000 kernel and/or grain length；
(d2) identify or aid in identification wheat seed mass of 1000 kernel；
(d3) identify or aid in identification wheat seed grain length；
(d4) kit with (d1) or (d2) or (d3) described purposes is prepared.
The present invention also protects specific DNA molecular (molecular labeling), as shown in the sequence 8 of sequence table.When N (also can use " n " table
Show) when being A, as high mass of 1000 kernel and/or the molecular labeling of long grain length.When N (also can use " n " to represent) is T as low thousand
The molecular labeling of weight and/or short grain length.The high mass of 1000 kernel is seed mass of 1000 kernel >=35g.The low mass of 1000 kernel is seed thousand
Weight ＜ 35g.The long grain length is seed grain length >=0.65mm.The short grain length is seed grain length ＜ 0.65mm.
The present invention also protects the primer sets I or the primer sets II.
The present invention also protects the applications of the primer sets I or the primer sets II, for following (d1) or (d2) or (d3) or
(d4)：
(d1) identify or assisting in and differentiate wheat seed character；The grain characters are mass of 1000 kernel and/or grain length；
(d2) identify or aid in identification wheat seed mass of 1000 kernel；
(d3) identify or aid in identification wheat seed grain length；
(d4) kit with (d1) or (d2) or (d3) described purposes is prepared.
The present invention also protects a kind of kit, contains the primer sets I or the primer sets II；The purposes of the kit
For following (d1) or (d2) or (d3)：
(d1) identify or assisting in and differentiate wheat seed character；The grain characters are mass of 1000 kernel and/or grain length；
(d2) identify or aid in identification wheat seed mass of 1000 kernel；
(d3) identify or aid in identification wheat seed grain length.
The present invention also protects any of the above methods described or specific DNA molecular or primer sets or kit in wheat breeding
Application.The purpose of the breeding to screen high mass of 1000 kernel wheat select based on 2144 SNP sites during the breeding
Genotype is the wheat of AA genotype.The purpose of the breeding carries out selecting base during the breeding to screen low mass of 1000 kernel wheat
In the wheat that the genotype of 2144 SNP sites is TT genotype.The purpose of the breeding carries out institute to screen long grain length wheat
Wheat of the genotype based on 2144 SNP sites for AA genotype is selected when stating breeding.The purpose of the breeding is the short grain of screening
Long wheat, carry out selecting wheat of the genotype based on 2144 SNP sites for TT genotype during the breeding.Described high thousand
The wheat that weight wheat is seed mass of 1000 kernel >=35g.The low mass of 1000 kernel wheat is seed mass of 1000 kernel ＜ 35g wheat.The length
Grain length wheat is seed grain length >=0.65mm wheat.The short grain length wheat is seed grain length ＜ 0.65mm wheat.
2144 SNP sites described in any of the above for sequence table sequence 8 from the nucleotides of 5 ' ends the 24th.
The present invention develops the SNP site related to wheat seed character, and devises base on the basis of the SNP site
In the primer sets of KASP technologies.Wheat seed character is identified using primer sets provided by the invention, it is quick and easy, accurate to have
The advantages that.The present invention is for wheat of the seed selection with different grain characters with fabulous application prospect.
Brief description of the drawings
A kind of original typing data in part of the step of Fig. 1 is embodiment 2.
Fig. 2 enters for the mass of 1000 kernel to various years improved variety and the variation tendency of the frequency of A/T allelic variations
The result of row analysis.
Embodiment
Following embodiment facilitates a better understanding of the present invention, but does not limit the present invention.Experiment in following embodiments
Method, it is conventional method unless otherwise specified.Test material used in following embodiments, it is certainly unless otherwise specified
What routine biochemistry reagent shop was commercially available.Quantitative test in following examples, it is respectively provided with and repeats to test three times, as a result make even
Average.
Embodiment 1, special SNP discovery and the design of special primer group
First, special SNP excavation
For trying wheat lines：Selection is distributed in China's difference area of wheat, 34 parts of wheat lines that grain characters differ greatly
(numbering C1-34, specific material information are shown in Table 1) the excavation material as pleomorphism site.
2nd, sequence alignment
It is each to carry out following steps operation respectively for examination wheat lines：
1st, genomic DNA of the extraction for examination wheat lines.
2nd, as template, the primer pair formed using TaTPP-F1 and TaTPP-R1 is carried out the genomic DNA extracted using step 1
PCR is expanded, and obtains pcr amplification product.
TaTPP-F1 (sequence 1 of sequence table)：5’-CGTGTGGTTGTTTGCGTG-3’；
TaTPP-R1 (sequence 2 of sequence table)：5’-CTAGATATAGGCGAGGGTTATTAC-3’.
3rd, the pcr amplification product for taking step 2 to obtain, cloning and sequencing is carried out.Every part of wheat lines survey 24 clones.
4th, sequence assembly and comparison are carried out.
24 cloning and sequencing results progress A genome sequences of every part of wheat lines are commented and connect and compare analysis, are found not
There are two kinds with the A genomic PCR amplification products for examination wheat.Two kinds of pcr amplification products are 2254bp, 5 ' ends with
TaTPP-F1 is consistent, and 3 ' ends are with TaTPP-R1 reverse complementals, and a kind of pcr amplification product is from 5 ' end 2121-2168 positions
Nucleotides is as shown in the sequence 3 of sequence table, and another pcr amplification product is from 5 ' end 2121-2168 positions nucleotides such as sequences
Shown in the sequence 4 of table
Based on the sequence alignment of all pcr amplification products for examination wheat, a SNP is found, is named as
2144SNP, it is that A/T is polymorphic.2144 SNP are the sequence 8 of sequence table from the nucleotides of 5 ' ends the 24th.
It is each to be shown in Table 1 for examination genotype of the wheat lines based on 2144 SNP.
5th, planted for examination wheat lines in Yuan Nei great solariums of Institute of Crop Science, Chinese Academy of Agricultural Science in October, 2012,
Normal irrigation fertilizing management, harvest seed in July, 2013 and measure mass of 1000 kernel.
Each mass of 1000 kernel for examination wheat lines is shown in Table 1.
Table 1
| Numbering | Title | Mass of 1000 kernel | Genotype | Numbering | Title | Mass of 1000 kernel | Genotype |
| C1 | Zhongyou9507 | 51.7g | AA | C18 | Shandong wheat 9 | 26.45g | TT |
| C2 | Zheng wheat 9023 | 44.1g | AA | C19 | Engrave virtuous 169 | 33.2g | TT |
| C3 | Climb 86001-3 | 52.8g | AA | C20 | Anhui 3 | 18.29g | TT |
| C4 | Shanxi wheat No. 8 | 41.3g | AA | C21 | Rob a wheat | 30.4g | TT |
| C5 | Laishou 953 | 42.05g | AA | C22 | Bai Dongmai | 15.75g | TT |
| C6 | Little Bai awns | 44.42g | AA | C23 | Orchid wheat | 28.6g | TT |
| C7 | Three cun | 53.66g | AA | C24 | Bai Mangmai | 29.85g | TT |
| C8 | Purple straw are red | 44.35g | AA | C25 | White flower wheat | 24.45g | TT |
| C9 | Red awns | 37.54g | AA | C26 | China spring | 27.35g | TT |
| C10 | Fish loach wheat | 44.29g | AA | C27 | Lv's drought 328 | 33.7g | TT |
| C11 | Shandong wheat 1 | 45.658g | AA | C28 | Agricultural university 139 | 32.05g | AA |
| C12 | Beijing 15 | 28.55g | TT | C29 | Jingyang 60 | 27.3g | TT |
| C13 | Shijiazhuang 54 | 33.28g | TT | C30 | Tobacco grower 15 | 34.05g | TT |
| C14 | Xuzhou 22 | 51.3g | AA | C31 | Bai Maizi | 24.45g | TT |
| C15 | Warm wheat No. 8 | 51.7g | AA | C32 | Fried dough twist plate | 20.9g | TT |
| C16 | Lankao 906 | 51.7g | AA | C33 | Red golden wheat | 23.4g | TT |
| C17 | It is short rich No. 3 | 34.464g | TT | C34 | March is yellow | 28.85g | TT |
34 in examination wheat, based on 2144 SNP genotype, 15 are AA genotype, and 19 are TT genotype.AA
The average mass of 1000 kernel of the seed for examination wheat of genotype is 45.91g, and the average mass of 1000 kernel of the seed for examination wheat of TT genotype is
27.54g。
Using mass of 1000 kernel be 35g as threshold value, wheat that seed mass of 1000 kernel is more than 35g is high mass of 1000 kernel wheat, seed thousand
The wheat less than 35g is low mass of 1000 kernel wheat again.If genotype of the wheat to be measured based on 2144 SNP is AA types, this is to be measured small
Wheat is the high mass of 1000 kernel wheat of candidate；If genotype of the wheat to be measured based on 2144 SNP is TT types, the wheat to be measured is time
The low mass of 1000 kernel wheat of choosing.Identify that above-mentioned 34 accuracys rate for the high mass of 1000 kernel wheat in examination wheat are 93% with this method
(14/15), identify that above-mentioned 34 accuracys rate for the low mass of 1000 kernel wheat in examination wheat are 100% (19/19) with this method.
2nd, the design of special primer group
The special SNP found according to step 1, it is as follows to design the primer sets based on KASP technologies：
2144F1 (sequence 5 of sequence table)：5’-
GAAGGTGACCAAGTTCATGCTTCACAGACTGCCACATCAGCGGCT-3’；
2144F2 (sequence 6 of sequence table)：5’-
GAAGGTCGGAGTCAACGGATTTCACAGACTGCCACATCAGCGGCA-3’；
2144C (sequence 7 of sequence table)：5’-TCTTGATAAATCAGTGCCAGGAG-3’；
3rd, the application of special primer group
The each of step 1 carries out following steps operation respectively for examination wheat lines：
1st, genomic DNA of the extraction for examination wheat lines.
2nd, for the genomic DNA extracted using step 1 as template, the special primer group designed using step 2 enters performing PCR amplification.
The reaction system of PCR amplifications：The μ L of template 2.2 (DNA content is about 100ng), MgCl2μ L of the aqueous solution 0.04,2 ×
μ L of Master Mix 2.5, the μ L of mix primer solution (mix primer solution contains 2144F1,2144F2 and 2144C) 0.056, use
ddH2O complements to 5 μ L.In PCR reaction systems, the concentration of magnesium ion is 0.2mM, and 2144F1 concentration is 12 μM, 2144F2
Concentration is 12 μM, 2144C concentration is 30 μM.2 × Master Mix full name is " KASP 2 × Master of V4.0 Mix
96/384 (Low Rox) ", Beijing LGC companies, catalog number KBS-1016-017.
The response procedures of PCR amplifications：94℃15min；(temperature of circulation is 65 for the first time by 95 DEG C of 20s, a certain temperature 20s
DEG C, each previous circulation of recycle ratio lowers 1 DEG C), 10 circulations；95 DEG C of 20s, 57 DEG C of 20s, 30 circulations.
PCR amplifications are run on the instruments of QuantStudio 7 of ABI companies production, export genotypic results automatically.Such as
The result of fruit gene parting is Alle 2 (blue round dot), is AA genotype for examination wheat；If the result of Genotyping is
Alle 1 (red spots), it is TT genotype for examination wheat；If genotypic results are Alle 1/Alle 2, for trying wheat
For AT genotype.
Each genotype call results for examination wheat are consistent with the genotype call results of step 1.
Embodiment 2, using special primer group detect large sample
It is each to be shown in Table 2 for examination wheat lines.
First, genotype detection
1st, genomic DNA of the extraction for examination wheat lines.
2nd, the genomic DNA extracted using step 1 as template, enter by the special primer group using two designs the step of embodiment 1
Performing PCR expands.
The reaction system of PCR amplifications：The μ L of template 2.2 (DNA content is about 100ng), MgCl2μ L of the aqueous solution 0.04,2 ×
μ L of Master Mix 2.5, the μ L of mix primer solution (mix primer solution contains 2144F1,2144F2 and 2144C) 0.056, use
ddH2O complements to 5 μ L.In PCR reaction systems, the concentration of magnesium ion is 0.2mM, and 2144F1 concentration is 12 μM, 2144F2
Concentration is 12 μM, 2144C concentration is 30 μM.2 × Master Mix full name is " KASP 2 × Master of V4.0 Mix
96/384 (Low Rox) ", Beijing LGC companies, catalog number KBS-1016-017.
The response procedures of PCR amplifications：94℃15min；(temperature of circulation is 65 for the first time by 95 DEG C of 20s, a certain temperature 20s
DEG C, each previous circulation of recycle ratio lowers 1 DEG C), 10 circulations；95 DEG C of 20s, 57 DEG C of 20s, 30 circulations.
PCR amplifications are run on the instruments of QuantStudio 7 of ABI companies production, export genotypic results automatically.Such as
The result of fruit gene parting is Alle 2 (blue round dot), is AA genotype for examination wheat；If the result of Genotyping is
Alle 1 (red spots), it is TT genotype for examination wheat；If genotypic results are Alle 1/Alle 2, for trying wheat
For AT genotype.
It is each to be shown in Table 2, table 3 and table 4 for examination genotype call results of the wheat lines based on 2144 SNP.Original point of part
Type data are shown in Fig. 1.
2nd, character detects
2002nd, 2005 and 2006, it will be planted for examination wheat lines in Luoyang, henan, and conventional water and fertilizer management, harvest seed
And it is wide (KW) to measure mass of 1000 kernel (TKW), grain length (KL) and grain.
The results are shown in Table 2 and (include each result for examination wheat, and the genotype for examination wheat lines of AA genotype
It is all for examination wheats average values).AT genotype for examination wheat lines to the results are shown in Table 3 (including each for examination wheat
As a result, and the genotype it is all for examination wheats average values).TT genotype the results are shown in Table 4 (bags for examination wheat lines
Include each result for examination wheat, and all average values for examination wheat of the genotype).From the point of view of main trend, AA genes
The mass of 1000 kernel of type wheat is more than TT genotype wheats, and the grain length of AA genotype wheats is more than TT genotype wheats.
Mass of 1000 kernel >=35g, it is defined as high mass of 1000 kernel；Mass of 1000 kernel ＜ 35g, are defined as low mass of 1000 kernel.Grain length >=0.65mm, it is fixed
Justice is long grain length；Grain length ＜ 0.65mm are defined as short grain length.The wheat of AA genotype is accredited as high mass of 1000 kernel wheat, long grain length
Wheat, accuracy rate the results are shown in Table 2.The wheat of TT genotype is accredited as low mass of 1000 kernel wheat, short grain length wheat, accuracy rate result
It is shown in Table 4.
Table 2
Table 3
Table 4
3rd, association analysis
For in examination wheat lines, the association analysis of Cultivars the results are shown in Table 5.As a result show：AA genotype supplies examination
Three annual mass of 1000 kernel of wheat are 41.50g, and the three annual mass of 1000 kernel for examination wheat of TT genotype are 36.45g, poor
It is different to reach the pole level of signifiance (P<0.01)；In grain length character, the wheat lines of the wheat lines of AA genotype compared with TT genotype
Seed is grown, and difference reaches the notable or pole level of signifiance (P<0.05 or P<0.01).It follows that for compared to TT genotype,
AA genotype is excellent grain characters genotype.
Table 5
Note：*P<0.05, * * P<0.01.
For in examination wheat lines, the association analysis of local varieties the results are shown in Table 6.As a result show：AA genotype supplies examination
Three annual mass of 1000 kernel of wheat are 38.9g, and the three annual mass of 1000 kernel for examination wheat of TT genotype are 31.55g, difference
Reach the pole level of signifiance (P<0.01)；In grain length character, the wheat lines seed of the wheat lines of AA genotype compared with TT genotype
Grain length, difference reach the notable or pole level of signifiance (P<0.05 or P<0.01).It follows that for compared to TT genotype, AA
Genotype is excellent grain characters genotype.
Table 6
Note：*P<0.05, * * P<0.01.
Embodiment 3,
Based on the result of embodiment 2, the frequency of mass of 1000 kernel and A/T allelic variations to various years improved variety
Variation tendency analyzed, as a result see Fig. 2.With the passage in age, increase is presented in the mass of 1000 kernel of China's Bred Wheat Varieties
Trend, consistent with this variation tendency, the frequencies of occurrences of the excellent allelic variation A in various years kind is also in rising
High trend, this shows that modern breeding has carried out very strong selection index system to the allelic variation, and the variation writes phase with grain representation
Close.Therefore, the mark can be as the functional label for improving Grain Weight in Common Wheat, carrying out improving yield of wheat molecular mark.
Claims (10)
1. a kind of identify or assisting in the method for differentiating wheat seed character, comprise the following steps：
It is AA genotype, AT genotype or TT to detect the genotype based on 2144SNP sites in the genomic DNA of wheat to be measured
Genotype；The grain characters of AA genotype wheats are better than TT genotype wheats；
The grain characters are excellent to be presented as that mass of 1000 kernel is high and/or grain length is grown；
The 2144SNP sites for sequence table sequence 8 from the nucleotides of 5 ' ends the 24th.
2. a kind of method identified or aid in identification wheat seed mass of 1000 kernel, comprises the following steps：
It is AA genotype, AT genotype or TT to detect the genotype based on 2144SNP sites in the genomic DNA of wheat to be measured
Genotype；It is high mass of 1000 kernel wheat if AA genotype, wheat candidate to be measured；If TT genotype, wheat candidate to be measured
For low mass of 1000 kernel wheat；
The high mass of 1000 kernel wheat is seed mass of 1000 kernel >=35g wheat；The low mass of 1000 kernel wheat is seed mass of 1000 kernel ＜ 35g
Wheat；
The 2144SNP sites for sequence table sequence 8 from the nucleotides of 5 ' ends the 24th.
3. a kind of method identified or aid in identification wheat seed grain length, comprises the following steps：
It is AA genotype, AT genotype or TT to detect the genotype based on 2144SNP sites in the genomic DNA of wheat to be measured
Genotype；It is long grain length wheat if AA genotype, wheat candidate to be measured；It is if TT genotype, wheat candidate to be measured
Short grain length wheat；
The long grain length wheat is seed grain length >=0.65mm wheat；The short grain length wheat is seed grain length ＜ 0.65mm's
Wheat；
The 2144SNP sites for sequence table sequence 8 from the nucleotides of 5 ' ends the 24th.
4. the application of the material of the genotype based on 2144SNP sites in the genomic DNA for detecting wheat, for following (d1)
Or (d2) or (d3) or (d4)：
(d1) identify or assisting in and differentiate wheat seed character；The grain characters are mass of 1000 kernel and/or grain length；
(d2) identify or aid in identification wheat seed mass of 1000 kernel；
(d3) identify or aid in identification wheat seed grain length；
(d4) kit with (d1) or (d2) or (d3) described purposes is prepared；
The 2144SNP sites for sequence table sequence 8 from the nucleotides of 5 ' ends the 24th.
5. specific DNA molecular, as shown in the sequence 8 of sequence table.
6. primer sets I, are made up of 2144F1,2144F2 and 2144C；
The primer 2 144F1 is following (b1) or (b2)：
(b1) single strand dna shown in the sequence 5 of sequence table；
(b2) have by sequence 5 by the substitution of one or several nucleotides and/or missing and/or addition and with sequence 5 identical
The DNA molecular of function；
The 2144F2 is following (b3) or (b4)：
(b3) single strand dna shown in the sequence 6 of sequence table；
(b4) have by sequence 6 by the substitution of one or several nucleotides and/or missing and/or addition and with sequence 6 identical
The DNA molecular of function；
The 2144C is following (b5) or (b6)：
(b5) single strand dna shown in the sequence 7 of sequence table；
(b6) have by sequence 7 by the substitution of one or several nucleotides and/or missing and/or addition and with sequence 7 identical
The DNA molecular of function.
7. primer sets II, are made up of TaTPP-F1 and TaTPP-R1；
The TaTPP-F1 is following (c1) or (c2)：
(c1) single strand dna shown in the sequence 1 of sequence table；
(c2) have by sequence 1 by the substitution of one or several nucleotides and/or missing and/or addition and with sequence 1 identical
The DNA molecular of function；
The TaTPP-R1 is following (c3) or (c4)：
(c3) single strand dna shown in the sequence 2 of sequence table；
(c4) have by sequence 2 by the substitution of one or several nucleotides and/or missing and/or addition and with sequence 2 identical
The DNA molecular of function.
8. the application of the primer sets of claim 6 or 7, for following (d1) or (d2) or (d3) or (d4)：
(d1) identify or assisting in and differentiate wheat seed character；The grain characters are mass of 1000 kernel and/or grain length；
(d2) identify or aid in identification wheat seed mass of 1000 kernel；
(d3) identify or aid in identification wheat seed grain length；
(d4) kit with (d1) or (d2) or (d3) described purposes is prepared.
9. a kind of kit, contain the primer sets of claim 6 or 7；The purposes of the kit is following (d1) or (d2)
Or (d3)：
(d1) identify or assisting in and differentiate wheat seed character；The grain characters are mass of 1000 kernel and/or grain length；
(d2) identify or aid in identification wheat seed mass of 1000 kernel；
(d3) identify or aid in identification wheat seed grain length.
10. in claims 1 to 3 described in specific DNA molecular described in any methods described or claim 5 or claim 6 or 7
Application of the kit described in primer sets or claim 9 in wheat breeding.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610757883.XA CN107794308B (en) | 2016-08-29 | 2016-08-29 | Specific SNP for identifying wheat grain traits and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610757883.XA CN107794308B (en) | 2016-08-29 | 2016-08-29 | Specific SNP for identifying wheat grain traits and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN107794308A true CN107794308A (en) | 2018-03-13 |
| CN107794308B CN107794308B (en) | 2020-05-12 |
Family
ID=61528015
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201610757883.XA Active CN107794308B (en) | 2016-08-29 | 2016-08-29 | Specific SNP for identifying wheat grain traits and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN107794308B (en) |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107794307A (en) * | 2016-08-29 | 2018-03-13 | 中国农业科学院作物科学研究所 | A kind of special SNP for identifying wheat seed character and its application |
| CN109022432A (en) * | 2018-09-04 | 2018-12-18 | 中国农业科学院作物科学研究所 | Identify the method and its primer special group of wheat tillering angle character |
| CN109468406A (en) * | 2018-12-28 | 2019-03-15 | 中国农业科学院作物科学研究所 | KASP label relevant to Wheat Seedling root system configuration and its application |
| CN111394506A (en) * | 2020-05-19 | 2020-07-10 | 黑龙江省农业科学院克山分院 | Wheat molecular marker and application thereof in identification of wheat grain weight character |
| CN112266975A (en) * | 2020-11-27 | 2021-01-26 | 山东省农业科学院作物研究所 | Primer group and kit for detecting KASP (Kaempferi-N-linked immunosorbent assay) marker related to POD (peroxidase) activity of wheat grains and application of primer group and kit |
| EP4111855A1 (en) | 2021-06-29 | 2023-01-04 | INIAV - Instituto Nacional de Invesigação Agrária E Veterinária, I.P. | Snp based panel for mediterranean wheat plant selection and breeding |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102086472A (en) * | 2009-12-08 | 2011-06-08 | 中国农业科学院作物科学研究所 | Gene relevant to thousand seed weight character of wheat, haplotype of gene and application thereof |
| CN104342484A (en) * | 2013-07-23 | 2015-02-11 | 中国农业科学院作物科学研究所 | Molecular marker related with wheat thousand grain weight and applications thereof |
| CN104805081A (en) * | 2015-04-30 | 2015-07-29 | 安徽农业大学 | Wheat grain heavy molecular marker and application thereof |
| CN107794307A (en) * | 2016-08-29 | 2018-03-13 | 中国农业科学院作物科学研究所 | A kind of special SNP for identifying wheat seed character and its application |
-
2016
- 2016-08-29 CN CN201610757883.XA patent/CN107794308B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102086472A (en) * | 2009-12-08 | 2011-06-08 | 中国农业科学院作物科学研究所 | Gene relevant to thousand seed weight character of wheat, haplotype of gene and application thereof |
| CN104342484A (en) * | 2013-07-23 | 2015-02-11 | 中国农业科学院作物科学研究所 | Molecular marker related with wheat thousand grain weight and applications thereof |
| CN104805081A (en) * | 2015-04-30 | 2015-07-29 | 安徽农业大学 | Wheat grain heavy molecular marker and application thereof |
| CN107794307A (en) * | 2016-08-29 | 2018-03-13 | 中国农业科学院作物科学研究所 | A kind of special SNP for identifying wheat seed character and its application |
Non-Patent Citations (3)
| Title |
|---|
| FENGMEI GAO等: ""Genome-Wide Linkage Mapping of QTL for Yield Components, Plant Height and Yield-Related Physiological Traits in the Chinese Wheat Cross Zhou 8425B/Chinese Spring"", 《FRONTIERS IN PLANT SCIENCE》 * |
| SHASHA WANG等: ""A Single-Nucleotide Polymorphism of TaGS5 Gene Revealed its Association with Kernel Weight in Chinese Bread Wheat"", 《FRONTIERS IN PLANT SCIENCE》 * |
| 刘胜男等: ""小麦RILs群体叶绿素含量和千粒重相关分析及QTL定位"", 《安徽农业大学学报》 * |
Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107794307A (en) * | 2016-08-29 | 2018-03-13 | 中国农业科学院作物科学研究所 | A kind of special SNP for identifying wheat seed character and its application |
| CN107794307B (en) * | 2016-08-29 | 2020-05-12 | 中国农业科学院作物科学研究所 | Specific SNP for identifying wheat grain traits and application thereof |
| CN109022432A (en) * | 2018-09-04 | 2018-12-18 | 中国农业科学院作物科学研究所 | Identify the method and its primer special group of wheat tillering angle character |
| CN109022432B (en) * | 2018-09-04 | 2020-06-09 | 中国农业科学院作物科学研究所 | Method for identifying wheat tillering angle character and special primer group thereof |
| CN109468406A (en) * | 2018-12-28 | 2019-03-15 | 中国农业科学院作物科学研究所 | KASP label relevant to Wheat Seedling root system configuration and its application |
| CN109468406B (en) * | 2018-12-28 | 2021-12-07 | 中国农业科学院作物科学研究所 | KASP marker related to wheat seedling stage root system configuration and application thereof |
| CN111394506A (en) * | 2020-05-19 | 2020-07-10 | 黑龙江省农业科学院克山分院 | Wheat molecular marker and application thereof in identification of wheat grain weight character |
| CN111394506B (en) * | 2020-05-19 | 2023-05-26 | 黑龙江省农业科学院克山分院 | Wheat molecular marker and application thereof in identifying wheat grain weight characteristics |
| CN112266975A (en) * | 2020-11-27 | 2021-01-26 | 山东省农业科学院作物研究所 | Primer group and kit for detecting KASP (Kaempferi-N-linked immunosorbent assay) marker related to POD (peroxidase) activity of wheat grains and application of primer group and kit |
| EP4111855A1 (en) | 2021-06-29 | 2023-01-04 | INIAV - Instituto Nacional de Invesigação Agrária E Veterinária, I.P. | Snp based panel for mediterranean wheat plant selection and breeding |
Also Published As
| Publication number | Publication date |
|---|---|
| CN107794308B (en) | 2020-05-12 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN107794308A (en) | Identify special SNP and its application of wheat seed character | |
| CN106676172B (en) | 212 SNP sites of tomato and its application in identification tomato variety authenticity and seed purity | |
| CN106636393B (en) | SNP marker and its application with pumpkin peel color gene linkage | |
| CN110305978A (en) | SNP site and its versatility molecular labeling, acquisition methods and application of a kind of and pepper fruit towards tight association | |
| CN107794307A (en) | A kind of special SNP for identifying wheat seed character and its application | |
| CN110295251A (en) | Chain SNP marker and its application with wheat available tillering QTL | |
| CN107475381A (en) | SNP marker and its application with tender flower stalk anthocyanidin gene linkage | |
| CN112159858A (en) | Molecular marker closely linked with purple cauliflower gene and application thereof | |
| CN105219858A (en) | Grain Weight in Common Wheat gene TaGS5-3A single nucleotide polymorphism and application thereof | |
| CN102277444A (en) | Method for quickly distinguishing grape varieties by random amplified polymorphic deoxyribonucleic acid (RAPD) | |
| CN106460063A (en) | SNP combination for Chinese cabbage germplasm resource diversity analysis and molecular breeding and application thereof | |
| CN107475414A (en) | A kind of SNP primer pairs of the method for screening the long high glycogen content parent shellfish of oyster and its correlation | |
| CN108531642B (en) | SSR molecular markers for identifying corn varieties and application thereof | |
| CN116790797A (en) | KASP primer group related to wheat grain weight and application thereof | |
| CN110283929A (en) | The relevant SNP marker 5-160 of capsicum epidemic disease resistant gene and its specific primer and application | |
| CN114164294B (en) | SNP locus related to green keeping property of Chinese cabbage and application thereof | |
| CN108841983A (en) | A kind of SSR primer of sugarcane overall length transcript profile data large-scale development | |
| CN105713983B (en) | A kind of molecular labeling and its application with rice Jiangnan evening neck blast resistance gene close linkage | |
| CN108517374A (en) | A kind of SNP marker and its application | |
| CN114480721A (en) | Method for identifying whether melon variety to be detected is thin-skin melon or thick-skin melon and special SNP primer combination thereof | |
| CN104762299B (en) | A kind of rice seedling resistant gene of salt qST2 and its molecule labelling method | |
| CN106434957B (en) | Application of the wheat molecular marker 4BL-699-1 and 4BL-699-2 in identification Plant Height in Wheat character | |
| CN108179222B (en) | Nucleotide sequence of chorismate mutase related to high yield of rice and application thereof | |
| CN107058601B (en) | SNP (Single nucleotide polymorphism) marker for identifying low stone cell content of pear pulp based on high-resolution dissolution curve and application of SNP marker | |
| CN105779581A (en) | Set of core SNP markers suitable for constructing variety nucleic acid fingerprint database of Chinese cabbage and applications of core SNP markers |
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 |