CN115852021A - SNP molecular marker for identifying wheat grain weight and grain length and application thereof - Google Patents
SNP molecular marker for identifying wheat grain weight and grain length and application thereof Download PDFInfo
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Abstract
The invention discloses an SNP molecular marker for identifying wheat grain weight and grain length and application thereof. The invention belongs to the field of biotechnology, and particularly comprises the steps of detecting polymorphism or genotype (allele) of an SNP locus in a wheat genome to be detected, and identifying or assisting in identifying the grain weight and the grain length of wheat according to the genotype, wherein the SNP locus is a locus on a wheat 2A chromosome, the nucleotide type of the SNP locus is G or T, and the SNP locus is 101 th nucleotide of a sequence 1 in a sequence table. The invention can be used for predicting the grain weight and grain length of wheat and breeding wheat. The substance for detecting the SNP site polymorphism and the genotype can be combined with other substances (such as substances for detecting the single nucleotide polymorphism or the genotype of other molecular markers related to the grain weight and the grain length of the wheat) to prepare the product for identifying the high grain weight and the grain length of the wheat.
Description
Technical Field
The invention belongs to the technical field of biology, and particularly relates to an SNP molecular marker for identifying wheat grain weight and grain length and application thereof.
Background
Wheat is an important food crop in China, the high yield and the stable yield of wheat are important guarantees of national food safety, the living standard of people in China and the national food safety are directly influenced by the high and low yield of wheat, and therefore high-yield breeding is a constant theme of wheat breeding in China. The yield constituent elements are relatively complex, particularly the number of ears and the number of grains per ear, the increase of thousand grain weight is relatively independent, the thousand grain weight of wheat is relatively stable, and the residual force reaches 59-80%. Zheng Tiancun considers that the contribution rate of each yield increase of Henan and Huanghuainan wheat is respectively thousand grain weight and grain number per ear and then ear number. Therefore, the thousand kernel weight plays a significant role in improving the yield per unit of wheat. The grain weight of wheat is mainly determined by the size of grains, and the size of the wheat grains can be further decomposed into the constituent elements such as grain length, grain width, grain thickness and the like. Wheat grain weight is mainly controlled by additive effect, the genetic force is up to 59-92%, and the wheat grain weight is quantitative character controlled by polygene. Research shows that the grain length and the grain width of the wheat are in extremely obvious positive correlation with the grain weight. Therefore, the research of controlling the genetic basis of the wheat grain size gene, cloning grain weight and grain shape related genes, exploring excellent allelic variation and developing a marker thereof by a modern molecular biology means have important significance for high yield breeding of wheat in China.
The gene positioning by using the parental genetic population or the natural population is an effective means for exploring new genes of the grain weight and the grain shape of the wheat. For example, giura et al, using wheat monosome studies, found that the 4A, 4B, 2B, 3A and 1B chromosomes were associated with grain length. Dholakia et al located QTLs associated with wheat grain length on 2BL, 2DL, 5BL, 6BS and 7 BL. Breseghello et al found by correlation analysis studies that 2D, 5A and 5B of wheat correlated with grain shape and grain weight, with Xwm539 correlated with grain length of wheat. Xwmc150b and Xbarc141 on chromosome 5A correlate with grain length and grain weight of wheat. Sun et al have QTL related to wheat grain length on chromosomes 1A, 1B, 2B, 4A and 4B, and xsws 24A-xsws 24c near the 4A chromosome are relatively stable in inheritance, and have a contribution rate to wheat grain length of 11.1-14.6%. Wang Ruixia et al used QTL mapping to detect a total of 17 grain-length-related QTLs under different environmental conditions, where two loci located on 2A and 2D were detectable under each environmental condition accounting for 10.7% and 14.7% of the phenotypic variation, respectively.
Although a great deal of QTL positioning work of wheat grain weight and grain shape genes is carried out by the prior people, the results of different researchers are poor in comparability due to the influence of factors such as mapping populations, genetic backgrounds, QTL mapping methods, marker types and the like. In addition, most QTLs still do not meet the requirement for molecular marker-assisted selection due to their small contribution rate to the grain weight and the grain shape phenotype and poor reproducibility between different years and environments. The invention takes a representative main-cultivated wheat variety (or an excellent variety) in Huang-Huai-Mai area as a test material, grain weight and grain length data are continuously investigated for 3 years in two environments of normal irrigation and drought, stable existing wheat grain weight and grain length related QTL are positioned through genome-wide association analysis (GWAS), KASP (competitive Allele-Specific PCR) molecular markers are designed according to SNP (Single nucleotide polymorphism) differences among different alleles for screening high-yield wheat germplasm materials, an effective detection means is provided for cultivating high-yield wheat varieties, and the invention has very important strategic significance for ensuring high and stable wheat yield and national grain safety and agricultural sustainable development.
Disclosure of Invention
The invention aims to solve the problem of how to identify the wheat grain weight and/or grain length in a high-throughput or auxiliary manner.
In order to solve the technical problems, the invention firstly provides a method for identifying or assisting in identifying wheat grain weight and/or grain length, which comprises the steps of detecting the genotype of an SNP locus in a wheat genome to be detected, and identifying or assisting in identifying the wheat grain weight and/or grain length according to the genotype, wherein the SNP locus is a locus on a wheat 2A chromosome, the nucleotide type of the SNP locus is G or T, and the SNP locus is the 101 th nucleotide of a sequence 1 in a sequence table.
The genotype is GG or TT, the GG is homozygous for the SNP with the nucleotide G, and the TT is homozygous for the SNP with the nucleotide T.
Taking a Chinese spring genome (IWGSC _ RefSeq _ v1.0, http:// 202.194.139.32/jbrown-1.12.3-release /) sequence of a wheat variety as a reference genome, wherein the SNP locus is a wheat 2A chromosome 608,865 and 127bp (specifically, 101 th locus of a sequence 1 in a sequence table).
As an embodiment, the method for identifying or assisting in identifying wheat grain weight and/or grain length may comprise the steps of:
(1) Using genome DNA of wheat to be detected as a template, and adopting a primer composition to carry out KASP marker detection; the primer composition consists of a primer A, a primer B and a primer C;
the primer A is a single-stranded DNA molecule with a nucleotide sequence of a sequence 2 in a sequence table or a single-stranded DNA with a nucleotide sequence of 22 th to 47 th sites of the sequence 2 in the sequence table;
the primer B is a single-stranded DNA molecule with a nucleotide sequence of sequence 3 in the sequence table or a single-stranded DNA with a nucleotide sequence of 22 th to 48 th sites of sequence 3 in the sequence table;
the primer C is a single-stranded DNA molecule with a nucleotide sequence of a sequence 4 in the sequence table;
(2) After the step (1) is finished, carrying out fluorescence detection to determine the genotype of the SNP of the wheat to be detected;
(3) And (3) identifying the grain weight and the grain length of the wheat to be detected according to the genotype result: the grain weight and/or grain length of the wheat to be detected with the SNP genotype of GG is superior to that of the wheat to be detected with the SNP genotype of TT.
The application of the method in wheat breeding also belongs to the protection scope of the invention.
The invention also provides the application of the substance for detecting the polymorphism or genotype of the SNP locus in the wheat genome in any one of the following parts:
(1) Identifying or assisting in identifying wheat grain weight and/or grain length;
(2) Breeding wheat;
(3) Preparing a product for identifying or assisting in identifying the grain weight and/or grain length of the wheat;
(4) Preparing a wheat breeding product;
the invention also provides a wheat breeding method.
The wheat breeding method provided by the invention comprises the steps of detecting the genotype of the SNP locus in a wheat genome, and selecting wheat with the genotype of GG as a parent for breeding, wherein the genotype of GG is homozygous type with the SNP of G.
As one implementation method, the method of wheat breeding may include the steps of:
(1) Taking the genome DNA of wheat to be detected as a template, and carrying out PCR amplification by adopting the primer group;
(2) After the step (1) is finished, carrying out fluorescence detection, and determining the genotype of the SNP locus of the wheat to be detected;
(3) And selecting the GG genotype wheat germplasm to breed the wheat with the advantages of long grain and heavy grain.
In the above method, the primer dissolution and preparation method may be: firstly, 3 primers are respectively added with ddH 2 Diluting O to 100 mu M, and preparing a primer working solution as follows: primer A12. Mu.L, primer B12. Mu.L, primer C30. Mu. L, ddH 2 O46. Mu.L was used as a KASP-labeled primer working solution and stored at-20 ℃ for further use.
In the above method, the reaction system of PCR may be: 1.5. Mu.L of template DNA, 0.0417. Mu.L of primer working solution, 0.75. Mu.L of 2 XKASP Master Mix (LGC Co., lot No. 13426773) and the reaction system was supplemented with sterile ultrapure water to 3. Mu.L.
In the above method, the PCR amplification can be performed on a high-throughput PCR instrument.
In the above method, the reaction procedure of PCR may be:
the first step is as follows: pre-denaturation at 94 ℃ for 15min;
the second step is that: denaturation at 94 deg.C for 20s, and renaturation for 20s (the first renaturation temperature is 61 deg.C, and the temperature is reduced by 0.6 deg.C per cycle) for 10 cycles; denaturation at 94 deg.C for 20s, and renaturation at 55 deg.C for 1min for 26 cycles;
the third step: extending at 72 deg.C for 3min, and storing at 4 deg.C.
In the above method, the method for determining the genotype of the SNP in the wheat to be detected may be: after the PCR reaction was completed, the reaction product was subjected to fluorescence data reading (data reading temperature of 40 ℃ or less) by converting the fluorescence signal into an analyzable value using a fluorescence signal reader (Omega) and a fluorescence detection system (Araya). The fluorescence scanning result is graphically displayed by utilizing an R software package, and the G base type has FAM fluorescence and is distributed near an x axis; the T base type has HEX fluorescence and is distributed near the y axis; samples with no detected signal are distributed near the origin.
The invention also provides a product for detecting the polymorphism or genotype of the SNP locus in the wheat genome.
The product for detecting the polymorphism or the genotype of the SNP locus in the wheat genome provided by the invention contains the substance for detecting the polymorphism or the genotype of the SNP locus in the wheat genome, and the product is any one of:
c1 Products for detecting single nucleotide polymorphisms or genotypes associated with wheat grain weight and/or grain length;
c2 Products that identify or assist in identifying wheat grain weight and/or grain length;
c3 Products for wheat breeding.
In the above applications, methods and products, the substance may be a reagent and/or an apparatus required for determining the polymorphism or genotype of the SNP site by at least one of the following methods: DNA sequencing, restriction enzyme fragment length polymorphism, single-strand conformation polymorphism, denaturing high performance liquid chromatography and SNP chip. The SNP chip comprises a chip based on nucleic acid hybridization reaction, a chip based on single base extension reaction, a chip based on allele-specific primer extension reaction, a chip based on one-step reaction, a chip based on primer connection reaction, a chip based on restriction enzyme reaction, a chip based on protein DNA binding reaction and a chip based on fluorescent molecule DNA binding reaction.
Optionally, the substance is D1), D2) or D3) as follows:
d1 The substance is a primer composition for amplifying a wheat genome DNA fragment including the SNP site;
d2 The substance is a PCR reagent containing the primer composition D1);
d3 The substance is a kit containing the primer composition D1) or the PCR reagent D2).
Alternatively, the amplification may be PCR amplification. The primer composition consists of the primer A, the primer B and the primer C.
D3 The kit may further comprise KASP Master Mix.
In the above applications, methods and products, the primer composition may or may not be labeled with a label. The label refers to any atom or molecule that can be used to provide a detectable effect and that can be attached to a nucleic acid. Labels include, but are not limited to, dyes; a radioactive label, such as 32P; binding moieties such as biotin (biotin); haptens such as Digoxin (DIG); a luminescent, phosphorescent, or fluorescent moiety; and a fluorescent dye alone or in combination with a portion of the emission spectrum that can be suppressed or shifted by Fluorescence Resonance Energy Transfer (FRET). Labels can provide signals detectable by fluorescence, radioactivity, colorimetry, gravimetry, X-ray diffraction or absorption, magnetism, enzymatic activity, and the like. Labels can be charged moieties (positive or negative) or alternatively, can be charge neutral. The label may comprise or be combined with a nucleic acid or protein sequence, provided that the sequence comprising the label is detectable. In some embodiments, the nucleic acid is detected directly (e.g., direct sequence read) without a label. The primer composition can be a primer composition consisting of a single-stranded DNA with a nucleotide sequence of 22 th to 47 th positions of a sequence 2 in a sequence table, a single-stranded DNA with a nucleotide sequence of 22 nd to 48 th positions of a sequence 3 in the sequence table and a single-stranded DNA with a nucleotide sequence of a sequence 4 in the sequence table, or a primer group consisting of a single-stranded DNA with a nucleotide sequence of 2 in the sequence table, a single-stranded DNA with a nucleotide sequence of 3 in the sequence table and a single-stranded DNA with a nucleotide sequence of 4 in the sequence table. The sequence 2 in the sequence table is composed of 47 nucleotides, the 1 st to 21 st nucleotides are FAM sequences (as markers), and the 22 nd to 47 th nucleotides are specific sequences; the sequence 3 in the sequence table is composed of 48 nucleotides, the 1 st to 21 st nucleotides are HEX sequences (as markers), and the 22 nd to 48 th nucleotides are specific sequences.
The invention also provides a DNA molecule, and the nucleotide sequence is shown as the sequence 1 in the sequence table.
The application of the DNA molecule is also within the protection scope of the invention. The application is specifically the application in any one of the following:
(1) Identifying or assisting in identifying wheat grain weight and/or grain length;
(2) Breeding wheat;
(3) Preparing a product for identifying or assisting in identifying the grain weight and/or grain length of the wheat;
(4) Preparing a wheat breeding product.
Optionally, in the above application, the DNA molecule serves as a detection target.
The substances for detecting the polymorphism and genotype of the SNP sites can be combined with other substances (such as substances for detecting single nucleotide polymorphism or genotype of other molecular markers related to the grain length and yield of wheat) to prepare the product for identifying the wheat variety with the grain weight and/or the grain length of the wheat.
Herein, the purpose of said breeding may comprise breeding high yielding wheat. The wheat may be a pure line or an inbred line.
The invention provides a primer composition and also provides a method for identifying or assisting in identifying wheat grain weight and grain length by using the primer composition. The method established by the invention can be used for predicting the grain weight and the grain length of the wheat, can be used for early screening of the wheat to be screened, can be used for wheat molecular marker-assisted breeding, and has important application value in the research of exploring wheat germplasm resources with increased grain length and yield and breeding wheat varieties with increased grain length and yield.
Drawings
FIG. 1 shows the KASP marker primer positions of the QTL locus QTGW.ICO-2A of common wheat grain weight and grain length in chromosome 2A for different allelic types. The SNP at the physical position 608,865,127bp (chip site AX-109290429) of wheat 2A chromosome is marked by shading, the KASP mark upstream primer position is marked by underlining, and the downstream primer position is marked by a box. The sequence in the figure is the sequence of the wheat 2A chromosome physical position 608,865,027bp-608,865, 226bp.
FIG. 2 is a diagram of wheat germplasm material Kasp _ QTGW2A marker detection result.
FIG. 3 is a box plot analysis result of thousand grain weight data of wheat germplasm of different allelic types of QTGW.
Fig. 4 is a result of boxplot analysis of grain length data of wheat germplasm with different allelic types of QTGW.
Detailed Description
The present invention is described in further detail below with reference to specific embodiments, which are given for the purpose of illustration only and are not intended to limit the scope of the invention. The examples provided below serve as a guide for further modifications by a person skilled in the art and do not constitute a limitation of the invention in any way.
The experimental procedures in the following examples, unless otherwise specified, were carried out in a conventional manner according to the techniques or conditions described in the literature in this field or according to the product instructions. Materials, reagents and the like used in the following examples are commercially available unless otherwise specified.
The quantitative experiments in the following examples, unless otherwise specified, were carried out in triplicate.
The wheat material in the following examples is of known variety and is described in the following references: <xnotran> ",,. . ,2015, 35 (1): 57-63.", ",,, ,,. () . , (2): 172-181.", ",,,. . , (3): 255-262.", ",,,,,. . , (): 31-39.", ", ,,,. . , (2): 196-202.", "Chen Shulin, cheng Xiyong, yu Kang, chang Xiangnan, bi Huihui, xu Haixia, wang Junsen, pei Xingxu, zhang Ziliang, zhan Kehui.Genome-wide association study of differences in 14agronomic traits under low-and high-density planting models based on the 660k SNP array for common wheat.Plant Breeding,2020,139 (2): 272-283.". </xnotran> The germplasm of common wheat is preserved by the wheat research center of the institute of grain and oil crops, academy of agriculture and forestry, hebei province, the public can obtain the biological material from the applicant, and the biological material is only used for repeating relevant experiments of the invention and cannot be used for other purposes.
Example 1 obtaining of region SNP marker chip site AX-109290429 and KASP marker primer set of QTGW.ICO-2A related to wheat grain weight and grain length
QTL location and discovery of chip site AX-109290429
The 15K SNP chip is utilized to carry out genotype typing on wheat varieties in Huang-Huai-Mai region, a whole genome correlation analysis method is adopted to carry out QTL positioning on the grain weight and the grain length of the wheat by combining with the phenotypic data of the grain weight and the grain length of the wheat varieties for many years, and the QTL for controlling the grain weight and the grain length of the wheat is positioned at the SNP locus AX-109290429, so that the QTL is converted into a KASP marker for molecular marker-assisted selective breeding. The SNP locus AX-109290429 is the 101 st position of the sequence 1, and the nucleotide type is G or T. K in the sequence 1 in the sequence table represents g or t.
Obtaining of primer set for Kasp _ QTGW2A labeled with KASP
Designing a primer group (namely KASP mark Kasp _ QTGW 2A) for detecting the SNP locus AX-109290429 based on the KASP technology, which is called KASP primer group for short. The KASP primer group consists of two upstream primers (primer A and primer B) and one downstream primer (primer C), and the specific sequences are shown in Table 1.
TABLE 1 KASP marker primer sequences for identifying common wheat QTGW.ICO-2A allelic variation
The primer A is a primer with a FAM fluorescent label sequence (base of an italic part) at the 5' end, the primer C is used for amplifying a fragment of which the SNP locus AX-109290429 is G, and a fluorescent signal of a FAM group can be read by a fluorescent signal reader;
the primer B is a primer with a HEX fluorescent label sequence (base of an italic part) at the 5' end, the primer C is used for amplifying a fragment with an SNP locus AX-109290429 as T, and a fluorescent signal of a HEX group can be read by a fluorescent signal reader.
Example 2 establishment of method for detecting SNP site AX-109290429 genotype by using KASP marker
The KASP mark Kasp _ QTGW2A is used for detecting different allele types of wheat grain weight and grain length QTGW. ICO-2A at the 2A chromosome physical position 608,865,127 (SNP locus AX-109290429).
PCR amplification System and program
Extracting genome DNA of common wheat leaf by CTAB method, adding 400 μ L TE for dissolving. The DNA is subjected to quality detection by 1% agarose gel electrophoresis, and the extracted DNA is required to have no obvious impurity, clear band and no degradation. After the concentration of the DNA is measured, the concentration is uniformly diluted to 28.3 ng/. Mu.L, and PCR amplification is carried out by taking the diluted wheat genome DNA as a template.
The primer sequences are shown in Table 1. Preparing KASP labeled primer working solution: firstly, 3 primers are respectively added with ddH 2 Diluting O to 100 mu M, and preparing a primer working solution according to the following formula: primer A12. Mu.L, primer B12. Mu.L, primer C30. Mu. L, ddH 2 O46. Mu.L. Storing at-20 deg.C for use.
The PCR amplification system is as follows: 1.5. Mu.L of template DNA, 0.0417. Mu.L of primer working solution, and 0.75. Mu.L of 2 XKASP Master Mix (LGC Co., lot No. 13426773), and the reaction system was supplemented with sterile ultrapure water to 3. Mu.L.
The PCR reaction program is: pre-denaturation at 94 ℃ for 15min; denaturation at 94 deg.C for 20s, and renaturation for 20s (the first renaturation temperature is 61 deg.C, and the temperature is reduced by 0.6 deg.C per cycle) for 10 cycles; denaturation at 94 deg.C for 20s, and renaturation at 55 deg.C for 1min for 26 cycles; extending at 72 deg.C for 3min, and storing at 4 deg.C.
The experiment was performed while setting a blank Control (CK) without adding template DNA to the reaction system, and setting 1 control for each plate.
2. Genotyping
After the PCR reaction is completed, the fluorescence signal is converted into analyzable values by a fluorescence signal reader (Omega) and a fluorescence detection system (Araya) to read the fluorescence data of the reaction product. The fluorescence scanning result is graphically displayed by utilizing an R software package, and the G base type (QTGW. ICO-2 Aa) has FAM fluorescence and is distributed near the x axis; the T base type (QTGW. ICO-2 Ab) has HEX fluorescence and is distributed near the y axis; samples with no detected signal are distributed near the origin.
The FAM excitation wavelength is 485nm, and the emission wavelength is 520nm. The HEX excitation wavelength is 535nm and the emission wavelength is 556nm. The excitation wavelength of the system reference fluorescence ROX is 575nm, and the emission wavelength is 610nm.
The result is shown in figure 2, and shows that only the fluorescent signal of HEX group exists, the genotype AX-109290429 of the wheat to be detected is TT (namely that the SNP locus AX-109290429 in the wheat genome is homozygotic of T); if the fluorescence signal only containing FAM group is displayed, the genotype of AX-109290429 of the wheat to be detected is GG (namely the SNP locus AX-109290429 in the wheat genome is homozygous for G).
Example 3 application of KASP marker Kasp _ QTGW2A in auxiliary identification of wheat grain weight and grain length in breeding
1.191 wheat germplasm weight and grain length phenotype typing
191 parts of wheat germplasm materials are planted on a test station on a levee of a research institute for grain and oil crops of agriculture and forestry academy of sciences in Hebei province in 2018-2019, 2019-2021 and 2020-2021 for 3 consecutive years. The design of completely random block groups is adopted, the row length is 3m, the row spacing is 30cm, and the plant spacing is 2cm. Set up normallyIrrigating (sowing in full soil, watering once in jointing stage and grouting stage, the irrigation quantity is 50m 3 Mu) and drought (sowing in sufficient soil moisture, no watering during the whole growth period). After the seeds are ripe and harvested, ten thousand-deep SC-G automatic seed test and a thousand-grain weight analyzer (ten thousand-deep detection technology) are used for measuring the grain length and the grain weight of the seeds, and a double-tail t-test method is adopted for carrying out statistical analysis on the grain length and the grain weight of each repeated group.
2. Identification of genotype of wheat SNP locus AX-109290429 by KASP marker
The genomic DNA of each test material was extracted and the genotype of the wheat tested was determined using the above molecular markers as described in example 2. The results are shown in Table 2 and FIG. 2. In FIG. 2, GG is a wheat with SNP locus AX-109290429 genotype of wheat material as GG, TT is a wheat with SNP locus AX-109290429 genotype of wheat material as TT, and CK is a blank control without adding template DNA in the reaction system.
Through KASP marker detection, 35 germplasms of 191 Chinese wheat germplasms are QTGW.ICO-2Aa allele type GG (namely SNP locus AX-109290429 genotype is GG), and 156 germplasms are QTGW.ICO-2Ab allele type TT (SNP locus AX-109290429 genotype is TT).
TABLE 2 wheat germplasm Kasp Label detection results and thousand grain weight (unit: g) data
Note: NA represents the missing data, and the wheat variety without the missing data is not subjected to the grain weight statistics.
TABLE 3 wheat germplasm Kasp mark detection result and grain length (unit: mm) data
Note: NA represents the missing data, and the wheat variety without the missing data is not subjected to the grain length statistics.
The data box plot analysis results of fig. 3, 4 are shown in conjunction with the statistical analysis results of tables 4 and 5: the grain weight and grain length mean values of the wheat germplasm with the SNP locus AX-109290429 genotype TT are lower than those of the wheat germplasm with the SNP locus AX-109290429 genotype GG in different years, and the grain weight and grain length mean values and the grain length mean values are remarkably or extremely remarkably different (P <0.05 or P < 0.01).
In breeding wheat with high grain weight and grain length, the wheat with SNP locus AX-109290429 genotype GG is preferably selected as a parent for breeding.
TABLE 4 statistical analysis results of the relationship between common wheat QTGW.ICO-2A allele types and thousand seed weights
Note: statistical analysis used a two-tailed t-test (P <0.05 for significant levels and P <0.01 for very significant levels).
TABLE 5 statistical analysis results of the relationship between the common wheat QTGW.ICO-2A allelic variation type and grain length
Note: statistical analysis used a two-tailed t-test (P <0.01 represents a very significant level of difference).
The present invention has been described in detail above. It will be apparent to those skilled in the art that the invention can be practiced in a wide range of equivalent parameters, concentrations, and conditions without departing from the spirit and scope of the invention and without undue experimentation. While the invention has been described with reference to specific embodiments, it will be appreciated that the invention can be further modified. In general, this application is intended to cover any variations, uses, or adaptations of the invention following, in general, the principles of the invention and including such departures from the present disclosure as come within known or customary practice within the art to which the invention pertains. The use of some of the essential features is possible within the scope of the claims attached below.
Claims (10)
1. The method for identifying or assisting in identifying the grain weight and/or grain length of wheat is characterized by comprising the following steps of: the method comprises the steps of detecting the genotype of an SNP locus in a wheat genome to be detected, and identifying or assisting in identifying the grain weight and/or grain length of wheat according to the genotype, wherein the SNP locus is a locus on a wheat 2A chromosome, the nucleotide type of the SNP locus is G or T, and the SNP locus is the 101 th nucleotide of a sequence 1 in a sequence table.
2. The application of the substance for detecting the polymorphism or genotype of the SNP locus in the wheat genome in any one of the following parts:
(1) Identifying or assisting in identifying the grain weight and/or grain length of wheat;
(2) Breeding wheat;
(3) Preparing a product for identifying or assisting in identifying the grain weight and/or grain length of the wheat;
(4) Preparing a wheat breeding product;
the SNP locus is a locus on a wheat 2A chromosome, the nucleotide type of the SNP locus is G or T, and the SNP locus is the 101 th nucleotide of a sequence 1 in a sequence table.
3. The method according to claim 1 or the use according to claim 2, characterized in that: the genotype of the SNP locus is GG or TT, the GG is homozygous for the SNP as G, and the TT is homozygous for the SNP as T; the grain weight and/or grain length of the wheat to be detected with the SNP genotype of GG is higher than that of the wheat to be detected with the SNP genotype of TT.
4. A method of wheat breeding characterized by: the method comprises detecting the genotype of the SNP in claim 1 in the genome of wheat, and selecting the wheat with the genotype of the SNP of GG as a parent for breeding, wherein the GG is a homozygous type of the SNP of G.
5. Use of the method of claim 1 or 4 in wheat breeding.
6. A product, characterized in that: the product containing the substance of claim 1, the product being any one of:
c1 Products for detecting single nucleotide polymorphisms or genotypes associated with wheat grain weight and/or grain length;
c2 Products that identify or assist in identifying wheat grain weight and/or grain length;
c3 Products for wheat breeding.
7. The use according to claim 2 or the product according to claim 6, characterized in that: the substances are D1), D2) or D3) as follows:
d1 The substance is a primer composition for amplifying a wheat genome DNA fragment including the SNP site;
d2 The substance is a PCR reagent containing the primer composition D1);
d3 The substance is a kit containing the primer composition D1) or the PCR reagent D2).
8. Use or product according to claim 7, characterized in that: the primer composition consists of a primer A, a primer B and a primer C;
the primer A is a single-stranded DNA molecule with a nucleotide sequence of a sequence 2 in a sequence table or a single-stranded DNA with a nucleotide sequence of 22 th to 47 th sites of the sequence 2 in the sequence table;
the primer B is a single-stranded DNA molecule with a nucleotide sequence of sequence 3 in the sequence table or a single-stranded DNA with a nucleotide sequence of 22 th to 48 th sites of sequence 3 in the sequence table;
the nucleotide sequence of the primer C is a single-stranded DNA molecule of a sequence 4 in a sequence table.
A dna molecule characterized by: the nucleotide sequence of the DNA molecule is sequence 1 in the sequence table.
10. The use of the DNA molecule of claim 9 in any one of,
(1) Identifying or assisting in identifying wheat grain weight and/or grain length;
(2) Breeding wheat;
(3) Preparing a product for identifying or assisting in identifying the grain weight and/or grain length of the wheat;
(4) Preparing a wheat breeding product.
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| CN117363785A (en) * | 2023-11-13 | 2024-01-09 | 河北省农林科学院粮油作物研究所 | Method for screening or assisting in screening wheat with different grain lengths and special primer group thereof |
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2022
- 2022-09-21 CN CN202211150104.1A patent/CN115852021A/en active Pending
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN117363785A (en) * | 2023-11-13 | 2024-01-09 | 河北省农林科学院粮油作物研究所 | Method for screening or assisting in screening wheat with different grain lengths and special primer group thereof |
| CN117363785B (en) * | 2023-11-13 | 2024-04-02 | 河北省农林科学院粮油作物研究所 | Method for screening or assisting in screening wheat with different grain lengths and special primer group thereof |
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