CN114622031B - SNP locus and CAPS molecular marker primer pair for identifying thousand seed weight characteristics of wheat and application of SNP locus and CAPS molecular marker primer pair - Google Patents
SNP locus and CAPS molecular marker primer pair for identifying thousand seed weight characteristics of wheat and application of SNP locus and CAPS molecular marker primer pair Download PDFInfo
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Abstract
The invention belongs to the field of bioengineering, and particularly relates to SNP loci and CAPS molecular marker primer pairs for identifying thousand seed weight characteristics of wheat and application thereof. The SNP locus provided by the invention is positioned at 1944bp of the coding region sequence of the wheat TaAGPL1-1B gene, has A/C base mutation, and has obvious correlation with the thousand-grain weight character of wheat. When the site is a C base, the wheat is of high thousand kernel weight. The SNP locus can be used for screening high thousand grain weight wheat, and is beneficial to wheat germplasm breeding.
Description
Technical Field
The invention belongs to the field of genetic engineering, and in particular relates to SNP loci and CAPS molecular marker primer pairs for identifying thousand seed weight characteristics of wheat and application thereof.
Background
Wheat (Triticum aestinum l.) genome is a heterologous hexaploid, consisting of A, B and D three very highly homologous genomes, which are large and complex, which provides challenges for development of wheat genetic breeding studies. Wheat is an important grain crop, and breeding of high-yield wheat varieties has been highly valued by breeders. Studies have shown that thousand kernel weight, which is primarily affected by kernel size traits, is a direct contributor to the most stable yield of wheat, and that per thousand kernel weight of wheat increases by 1g, the yield of wheat per hectare increases by 140-160kg (Tian et al 2006). It can be seen that increasing thousand kernel weight of wheat to increase yield is one of the most important breeding goals for wheat. The wheat yield is composed of three elements of small spike number, spike number and thousand grain weight, and the improvement of the Chinese wheat yield mainly depends on the increase of spike number and thousand grain weight (Wang Lanfen, etc.); and the starch accounts for 65-75% of the dry weight of the wheat grains, and the synthetic amount and the proportion of the starch obviously influence the thousand grain weight of the wheat, thereby influencing the yield of the wheat.
Starch synthesis is a complex process, and six enzymes involved in endosperm starch synthesis have been identified, including adenosine diphosphate glucose pyrophosphorylase (AGPase), particle-bound starch synthase (GBSS), soluble Starch Synthase (SS), branching Enzyme (BE), debranching enzyme (DBE), and glucose-6 phosphotransferase (GPT 1). Because of the interactions of these enzymes, it is ensured that plants synthesize new starch to meet their own needs, where AGPase catalyzes the formation of glucose 1-phosphate (G-1-P) and Adenosine Triphosphate (ATP) in higher plants (ADPG), which is a substrate for amylose and amylopectin synthesis, and its concentration directly affects the rate and efficiency of starch synthesis, and AGPase is a key enzyme in starch synthesis pathways in higher plants such as wheat, rice and maize. Wang Yuefu et al showed that the change in AGPase activity was significantly positively correlated with the accumulation of grain filling rate and starch content. The corn AGPase gene containing specific site mutation is transferred into wheat by Smitansky et al, so that the weight of seeds and the starch content of the seeds can be obviously increased; however, if the gene is mutated or artificially knocked out, the activity of the enzyme is reduced or eliminated, thereby inhibiting starch synthesis. So far, there is little report on the screening of excellent allelic variation of TaAGPS gene and its molecular marker in wheat, which greatly hinders the application of wheat starch synthesis key gene AGPase in improving wheat yield traits. Therefore, the gene TaAGPL1 related to the thousand seed weight of the wheat is cloned, related functional markers are developed, haplotype analysis is carried out, and correlation analysis is carried out on the haplotype and the thousand seed weight characteristics to find excellent haplotypes, so that the method has extremely important significance for improving the thousand seed weight of the wheat and obtaining new varieties of high-yield wheat.
Disclosure of Invention
The invention aims to provide a SNP locus and CAPS molecular marker primer pair for identifying the thousand-grain weight character of wheat and application thereof, detect excellent allelic variation of the TaAGPL1-1B gene of the wheat, screen and identify the thousand-grain weight character of the wheat and improve the breeding efficiency of the germplasm of the wheat with high thousand-grain weight.
The invention provides an SNP locus for identifying thousand-grain weight characters of wheat, wherein the SNP locus is positioned at 1944bp of a coding region sequence of a TaAGPL1-1B gene of the wheat, and polymorphism is A/C.
Preferably, the CDS sequence of the coding region of the TaAGPL1-1B gene is shown as SEQ ID No. 1.
The invention also provides a CAPS molecular marker primer pair for identifying the SNP locus, which comprises an upstream primer CAPS-F with a sequence shown as SEQ ID NO.2 and a downstream primer CAPS-R with a sequence shown as SEQ ID NO. 3.
The invention also provides a method for detecting excellent allelic variation of TaAGPL1-1B gene in wheat, which comprises the following steps:
carrying out PCR amplification on genome DNA of the material to be detected by utilizing the primer to obtain a PCR amplification product;
carrying out enzyme digestion on the PCR amplification product by using restriction enzyme Taq I, and detecting polymorphism of the enzyme digestion product;
when the enzyme digestion product comprises two short fragments of 401bp and 42bp, the allelic variation type is TaAGPL1-1B-C; when the cleavage product has no short fragment, the allelic variation type is TaAGPL1-1B-A.
Preferably, the reaction system used for PCR amplification comprises, in terms of 50. Mu.L: 20ng of genomic DNA of the material to be tested, 10. Mu.M of upstream primer CAPS-F0.3. Mu.L, 10. Mu.M of downstream primer CAPS-R0.3. Mu.L, 2X KOD OneTM PCR Master Mix. Mu.L and the balance of ddH 2 O；
The PCR amplification reaction is carried out by the following steps: the first stage is pre-denaturation at 94 ℃ for 3min; the second stage of denaturation at 98 ℃ for 10s, annealing at 59 ℃ for 30s, extension at 68 ℃ for 3min and 32 cycles; and a third stage: final extension at 68℃for 5min.
The invention also provides an application of the SNP locus or the CAPS molecular marker primer pair or the method in wheat breeding.
The invention also provides an application of the SNP locus or the CAPS molecular marker primer pair or the method in wheat auxiliary breeding.
The invention also provides an application of the SNP locus or the CAPS molecular marker primer pair or the method in identifying the thousand seed weight type of wheat.
The invention also provides application of the SNP locus or the CAPS molecular marker primer pair or the method in screening high thousand grain weight wheat.
The invention also provides a method for breeding high thousand grain weight wheat with the assistance of molecular markers, which utilizes the CAPS molecular marker primer pair to carry out molecular markers on target wheat, screens the 1944bp position of the coding region sequence of the wheat TaAGPL1-1B gene, and carries out subsequent screening on germplasm with polymorphism C.
The SNP locus provided by the invention is positioned at 1944bp of a coding region of a wheat TaAGPL1-1B gene, and the polymorphism is A/C; when the base is C, the wheat is high in thousand grain weight. The SNP locus can be used for distinguishing high thousand grain weight wheat from low thousand grain weight wheat.
The invention adopts the CASP molecular marker primer pair to detect the allelic variation of the TaAGPL1-1B gene related to thousand seed weight in wheat, and can finish the identification of the sample by simple DNA extraction, PCR specific amplification and enzymolysis detection, thereby being convenient for detecting and screening wheat varieties or strains with high thousand seed weight and greatly accelerating the breeding process of wheat high-yield varieties.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings that are required to be used in the embodiments will be briefly described below.
FIG. 1 shows the results of PCR amplification of the coding region of TaAGPL1-1B with a promoter;
FIG. 2 shows the mutation sites and two haplotypes of TaAGPL1-1B promoter and coding region sequence in common hexaploid wheat;
FIG. 3 shows the nucleotide sequence differences of two wheat types Shan Bei of TaAGPL1-1B-Hap1 and TaAGPL1-1B-Hap2 in the coding region (1 bp-3351 bp) of the TaAGPL1-1B gene;
FIG. 4 shows the nucleotide sequence differences of the promoter regions (-2225 bp-1 bp) of two wheat haplotypes TaAGPL1-1B gene of TaAGPL1-1B-Hap1 and TaAGPL1-1B-Hap 2;
FIG. 5 shows thousand kernel weight analysis results of TaAGPL1-1B in two haplotype wheat varieties;
FIG. 6 shows the nucleotide sequence differences of two wheat haplotypes of TaAGPL1-1B-Hap1 and TaAGPL1-1B-Hap2 for amplifying the fragment of coding region of TaAGPL1-1B gene (1902 bp-2344 bp);
FIG. 7 shows the result of electrophoresis detection of the TaAGPL1-1B promoter haplotype cleavage product.
Detailed Description
The invention provides an SNP locus for identifying thousand seed weight characters of wheat, which is positioned at 1944bp of a coding region sequence of a wheat TaAGPL1-1B gene, and has polymorphism A/C; when the base is C, the wheat is thousand-grain weight wheat.
In the invention, the SNP locus is positioned at 1944bp locus of the coding region sequence of the wheat TaAGPL1-1B gene, and the locus has A/C base mutation and has obvious correlation with the thousand-grain weight character of wheat. The nucleotide sequence of the coding region of the TaAGPL1-1B gene is shown as SEQ ID No.13 or SEQ ID No. 14; the CDS sequence of the coding region of the TaAGPL1-1B gene is shown as SEQ ID No. 1. The polymorphism site of the SNP is positioned at 1944bp of the coding region of the TaAGPL1-1B gene, and the polymorphism is A/C. The thousand seed weight performance of the wheat to be detected can be determined by identifying and typing the SNP locus: when the locus is C base, the thousand grain weight character of the wheat is high thousand grain weight.
The invention provides a CAPS molecular marker primer pair for identifying SNP loci, which comprises an upstream primer CAPS-F (5'-CATGTGCAGAAAGTGGACA-3') with a sequence shown as SEQ ID NO.2 and a downstream primer CAPS-R (5'-AGAAGGATCTGATTGTTCCA-3') with a sequence shown as SEQ ID NO. 3. The CAPS molecular marker primer pair can detect allelic variation of TaAGPL1-1B genes related to thousand kernel weight in wheat, detect and screen wheat varieties or strains with high thousand kernel weight, and greatly accelerate the breeding process of wheat high-yield varieties.
The invention provides a method for detecting allelic variation of TaAGPL1-1B gene in wheat by using the CAPS molecular marker primer pair, which comprises the following steps:
carrying out PCR amplification on genome DNA of the material to be detected by utilizing the primer pair to obtain a PCR amplification product;
carrying out enzyme digestion on the PCR amplification product by using restriction enzyme Taq I, and detecting polymorphism of the enzyme digestion product;
when the enzyme digestion product comprises two short fragments of 401bp and 42bp, the allelic variation type is TaAGPL1-1B-C; when the cleavage product has no short fragment, the allelic variation type is TaAGPL1-1B-A.
The invention firstly extracts the genome DNA of the material to be detected. The material to be tested according to the invention is preferably wheat. The invention has no special requirements on the types of the wheat, and any types of wheat can be used. The method for extracting the genome DNA of the wheat to be detected is not particularly limited, and the method for extracting the genome of the plant cells is conventional in the art. In the specific implementation process of the invention, the wheat leaves or other tissues to be detected are taken as materials, and the CTAB method is used for extraction. The CTAB method of the invention is carried out according to a conventional method.
After obtaining the wheat genome DNA, the invention uses the wheat genome DNA to be detected as a template, and uses the primer pair to carry out PCR amplification reaction to obtain a PCR amplification product. The reaction system used in the PCR amplification reaction of the present invention preferably comprises, in terms of 50. Mu.L: 200ng of wheat genome DNA, 10. Mu.M of upstream primer CAPS-F0.3. Mu.L, 10. Mu.M of downstream primer CAPS-R0.3. Mu.L, 2X KOD OneTM PCR MasterMix. Mu.L and the balance of ddH 2 O. The KOD OneTM PCR Master Mix of the present invention was purchased from Toyo-spinning biosystems. The concentration of the wheat genome DNA is preferably 100-200 ng/. Mu.L; OD of the genomic DNA 260 /OD 280 The ratio is preferably 0.6 to 0.8.
The procedure for the PCR amplification reaction according to the invention is preferably: the first stage is pre-denaturation at 94 ℃ for 3min; the second stage of denaturation at 98 ℃ for 10s, annealing at 59 ℃ for 30s, extension at 68 ℃ for 3min and 32 cycles; and a third stage: final extension at 68℃for 5min.
After the completion of the PCR amplification, the PCR amplification product is preferably stored at 4 ℃. The primer pair can be used for amplifying TaAGPL1-1B fragments (1901 bp-2344 bp) in different haplotypes of wheat, and a PCR amplification product obtained by the PCR amplification reaction contains a DNA fragment with target SNPs sites.
After the PCR amplification product is obtained, the amplification product is recovered and the concentration is measured. The invention uses restriction enzyme Taq I to carry out enzyme digestion on the PCR amplified product to obtain an enzyme digestion product. The reaction system used for the cleavage of the present invention preferably comprises, in terms of 10. Mu.L: PCR amplification product 300ng, taq I0.5. Mu.L, buffer 1. Mu. L, BSA 1. Mu.L, ddH 2 O to 10. Mu.L. The Taq I restriction enzyme of the invention is purchased from da Lian Bao Bio Inc.
After the enzyme digestion product is obtained, the polymorphism of the enzyme digestion product is detected, and the allelic variation type of the TaAGPL1-1B gene in the wheat is judged according to the detection result. The assay of the invention is preferably performed by agarose gel electrophoresis. The invention preferably detects allelic variation of TaAGPL1-1B gene in wheat according to fragment size of enzyme cutting product, and can judge thousand grain weight character of wheat by allelic variation type. Specifically, when the enzyme digestion product comprises two short fragments of 401bp and 42bp, the allelic variation type is TaAGPL1-1B-C, and the wheat with high thousand grain weight is identified; when the cleavage product has no short fragment, the allelic variation type is TaAGPL1-1B-A, and the low thousand grain weight wheat is identified.
The invention also provides application of the SNP locus or CAPS molecular marker primer pair or method in any one of (a) to (d):
(a) Wheat breeding; (b) wheat assisted breeding; (c) identifying the thousand kernel weight type of wheat; (d) screening high thousand kernel weight wheat.
The invention also provides a method for breeding high thousand grain weight wheat with the assistance of molecular markers, which utilizes the CAPS molecular marker primer pair to carry out molecular markers on target wheat, screens the 1944bp position of the coding region sequence of the wheat TaAGPL1-1B gene, and carries out subsequent screening on germplasm with polymorphism C.
The technical solutions provided by the present invention are described in detail below with reference to the drawings and examples for further illustrating the present invention, but they should not be construed as limiting the scope of the present invention.
Example 1
1. Primer design
Based on the sequence differences of the three copies 1A, 1B, 1D of the TaAGPL1 gene in the wheat genome database (http:// plants. Ensemb. Org/Triticum_aestium/Info/Index), taAGPL1-1B specific Promoter amplification primers TaAGPL1-1B-Promoter-SF (SEQ ID No. 4) and TaAGPL1-1B-Promoter-SR (SEQ ID No. 5) were designed, and the coding region amplification primers TaAGPL1-1B-CDS-SF1 (SEQ ID No. 6), taAGPL1-1B-CDS-SR1 (SEQ ID No. 7), taAGPL1-1B-CDS-SF2 (SEQ ID No. 8) and TaAGPL1-1B-CDS-SR2 (SEQ ID No. 9) were completed by Henan Shang Ya biotechnology company, the specific nucleotide sequences were as follows:
TaAGPL1-1B-Promoter-SF：5’-TCGCTCGCTCTTCTTCAGC-3’；
TaAGPL1-1B-Promoter-SR：5’-TGCCACCGCCGAGTATGACG-3’；
TaAGPL1-1B-CDS-SF1：5’-ATGCTTCTCAACTTCTCGT-3’；
TaAGPL1-1B-CDS-SR1：5’-GCTGAGACGAGTGCTAACC-3’；
TaAGPL1-1B-CDS-SF2：5’-CATGTGGATGACAATGCTGAC-3’；
TaAGPL1-1B-CDS-SR2：5’-CTTACTACTGATGCTGCCTGC-3’。
2. extraction of total DNA from wheat
321 common wheat varieties are randomly selected, and total DNA of wheat is extracted by adopting a CTAB method: first, wheat leaves or other tissues were flash-ground with liquid nitrogen, and 1mL of the preheated CTAB extract was added, followed by a water bath at 65℃for 1h. Then, after centrifugation at 12000g, the supernatant was taken and added with an equal volume of chloroform: the isoamyl alcohol (24:1) mixture was shaken. Centrifuge 12000g for 10min, transfer the supernatant to a new tube, add 2 volumes of absolute ethanol, mix well, stand at-20 ℃ for 30min. The supernatant was centrifuged again, washed twice with 75% ethanol, dried in the air, and 100. Mu.L of sterile water was added to dissolve the DNA.
Amplification and sequencing of the TaAGPL1-1B Gene promoter and coding region
Taking 2321 hexaploid wheat DNA as a template, respectively utilizing three pairs of primers of TaAGPL1-1B-Promoter-SF/R, taAGPL1-1B-CDS-SF1/R1 and TaAGPL1-1B-CDS-SF2/R2, and amplifying the sequence of the Promoter and the coding region CDS of TaAGPL1-1B by using high-fidelity PCR enzyme KOD OneTM PCR Master Mix (purchased from Toyobo Co., ltd.), wherein the nucleotide sequences corresponding to the Promoter and the coding region CDS of TaAGPL1-1B are shown as SEQ ID No.10 and SEQ ID No.1 respectively.
The PCR system comprises: 25 mu L KOD OneTM PCR Master Mix (2X), 0.3 mu L (10 mu M) of each of the upstream and downstream primers, 200ng of DNA template, and ddH supplementation 2 O to a total volume of 50. Mu.L.
The PCR reaction procedure was: in the first stage, pre-denaturation is carried out for 3min at 94 ℃; a second stage, denaturation at 98 ℃ for 10s, annealing at 59 ℃ for 30s, extension at 68 ℃ for 3min,32 cycles; in the third stage, the temperature is kept at 68 ℃ for 5min.
The amplification results are shown in FIG. 1: wherein (a) in FIG. 1 is a Promoter fragment of TaAGPL1-1B amplified by using TaAGPL1-1B-Promoter-SF/R as an amplification primer pair, and the size is 2320bp; in FIG. 1, (B) is a fragment of coding region of TaAGPL1-1B amplified by using TaAGPL1-1B-CDS-SF1/R1 as an amplification primer pair, the fragment having a size of 1873bp; in FIG. 1, (c) is a fragment of the coding region of TaAGPL1-1B amplified by using TaAGPL1-1B-CDS-SF2/R2 as an amplification primer pair, and the size is 1964bp.
The PCR amplified target fragments of 321 varieties are respectively recovered and connected with ZT4-Blunt cloning vector (purchased from the allied biological company) to transform escherichia coli, more than three positive clones are selected from each wheat variety, and the sequence is sequenced by using universal primers T7-F/R (purchased from the allied biological company, the nucleotide sequence of the upstream primer T7-F of T7 is shown as SEQ ID No. 11: 5'-TAATACGACTCACTATAGGG-3', and the nucleotide sequence of the downstream primer T7-R of T7 is shown as SEQ ID No. 12: 5'-AAGAACATCGATTTTCCATGGCAG-3').
Sequence alignment and haplotype analysis of TaAGPL1-1B Gene promoter and coding region
The sequencing results were spliced and aligned using DNAMAN software (http:// www.lynnon.com). Sequence alignment results show that 4 SNPs (1944A/C, 2209G/C,2566C/T and 2209G/A) exist in the TaAGPL1-1B coding region (1 bp-3351 bp) in 321 wheat varieties, wherein 2 SNPs (1944A/C and 2566C/T) are located on exons, but do not cause the change of coding amino acids, and belong to nonsense mutation sites.
8 SNP polymorphism mutation sites exist in a promoter region (-2225 bp-1 bp), haplotype identification is carried out on the promoter by DnaSP 5.10 software (http:// www.ub.edu/DnaSP), and the result shows that the 12 SNPs are tightly linked to form two haplotypes, which are named as TaAGPL1-1B-Hap1 and TaAGPL1-1B-Hap2 respectively, and the nucleotide sequence differences of the two haplotypes of TaAGPL1-1B-Hap1 and TaAGPL1-1B-Hap2 in a coding region (1 bp-3351 bp) of a TaAGPL1-1B gene are shown in figure 3; the nucleotide sequence differences in the promoter region (-2225 bp-1 bp) of the TaAGPL1-1B gene are shown in FIG. 4.
5. Thousand kernel weight analysis of different haplotype wheat varieties in hexaploid wheat
Step 1321 the haplotypes of wheat varieties and the thousand kernel weights in hexaploid wheat are shown in Table 1, and the average thousand kernel weights of TaAGPL1-1B-Hap1 haplotypes and the average thousand kernel weights of TaAGPL1-1B-Hap2 haplotypes are compared as shown in Table 2 and FIG. 5.
TABLE 1 haplotypes and thousand kernel weights of different wheat varieties
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TABLE 2 average thousand kernel weight of hexaploid wheat TaAGPL1-1B haplotypes
As can be seen from tables 1 to 2 and FIG. 5, of 321 wheat varieties, 166 varieties of wheat were in total Hap1 haplotype, and the average thousand grain weight was 34.91g;155 varieties of wheat are of the Hap2 haplotype, the average thousand grain weight is 42.20g, the average thousand grain weight of the wheat variety of the Hap2 haplotype is remarkably higher than that of the wheat variety of the Hap1 haplotype, and the wheat variety of the Hap2 is of the high thousand grain weight excellent haplotype.
CAPS molecular marker development
As can be seen from the description of FIG. 2, there is a single base difference between the two haplotypes TaAGPL1-1B, one of the SNP in Hap 2: 1944bp (C/A) was located in the sequence TCGA; one of the SNPs of Hap 1: 1944bp (C/A) was located in the sequence TAGA, and the differential amplification polymorphic (CAPS) molecular markers herein were developed and designated CAPS-1944 as the polymorphic marker in the 1944bp sequence of the coding region of the TaAGPL1-1B gene.
Primers CAPS-F and CAPS-R are designed at the upstream and downstream of the SNP for identifying CAPS-1944, the step 1 is used for preparing the Hap1 haplotype wheat variety Hap1 wheat-two mango, the Hap1 haplotype wheat-violet 13, the Hap1 haplotype wheat-five-flower head, the Hap1 haplotype wheat-Scutellaria wheat, the Hap1 haplotype wheat-Kenong 199 and the Hap1 haplotype wheat-stone 4185, and the DNA of Hap2 haplotype wheat variety Hap2 haplotype wheat as original wheat No.9, hap2 haplotype wheat ocean Bai Mai, hap2 haplotype wheat crop 9204, hap2 haplotype wheat small elytrigia 54, hap2 haplotype wheat Beijing 411 and Hap2 haplotype wheat stone 828 as template, and the nucleotide sequence difference of TaAGPL1-1B fragment (1902 bp-2344 bp), taAGPL1-1B-Hap1 and TaAGPL1-1B-Hap2 two wheat haplotype amplified TaAGPL1-1B gene coding region fragment (1902 bp-2344 bp) in different wheat varieties is shown in figure 6. The purified PCR product is subjected to enzyme digestion by using Taq I, the enzymolysis product is subjected to electrophoresis inspection, and the electrophoresis detection result is shown in figure 7, wherein the figure 7 sequentially shows that Hap1 haplotype wheat two-mango, hap1 haplotype wheat-violet 13, hap1 haplotype wheat five-flower head, hap1 haplotype wheat-cotton seed, hap1 haplotype wheat-wheat farm 199, hap1 haplotype wheat-stone 4185, hap2 haplotype wheat-winding original selection No.9, hap2 haplotype wheat-ocean Bai Mai, hap2 haplotype wheat-wheat farm 9204, hap2 haplotype wheat-elytrigia 54, hap2 haplotype wheat-stone 411 and Hap2 haplotype wheat-stone new 828 from left to right.
As can be seen from FIG. 7, after enzyme digestion, the amplified product of the Hap1 haplotype wheat variety is still a single 443bp band, while the amplified product of the Hap2 haplotype material is changed into two fragments, namely 401bp and 42bp, after enzyme digestion, the CAPS-1944 molecular marker can distinguish the Hap2 from other haplotypes, has obvious correlation with the thousand grain weight shape of wheat, can screen high thousand grain weight wheat by utilizing the SNP locus, and can be applied to molecular design breeding of new high thousand grain weight wheat varieties.
Although the foregoing embodiments have been described in some, but not all, embodiments of the invention, it should be understood that other embodiments may be devised in accordance with the present embodiments without departing from the spirit and scope of the invention.
Sequence table information:
DTD version v1_3
File name-11569C 23FI10216-xulie. Xml
Software name WIPO Sequence
Software version 2.3.0
Date of production 2023-08-24
Basic information:
current application/intellectual property office CN
Current application/number 202210269669.5
Current application/filing date 2022-03-18
Current application/applicant archive name 11569C23FI10216
Applicant name or Henan agricultural university
Applicant name or name/language zh
Applicant name or name/latin name Henan Agricultural University
The names of the inventors are Kang Guozhang, meng, wang Pengfei, ge Jiang, guo Tiancai, liu Guoqin, yang Xitang, peng Yanyan
Inventor name/language zh
The name of the inventor/Latin is Kang Guozhan, meng Yanjun, wang Pengfei, ge Qiang, guo Tiancai, liu Guoqin, yang Xitang, peng Yang an
The invention discloses a SNP locus and CAPS molecular marker primer pair for identifying thousand grain weight characteristics of wheat and application thereof (zh)
Total sequence 14
Sequence:
serial number (ID) 1
Length 1569
Molecular type DNA
Feature location/qualifier:
- source, 1..1569
> mol_type, other DNA
> organism, synthetic construct
residues:
atgtcatcga tgcagttcag cagcgtgctg cccctggagg gcaaagcgtg catctccccc 60
gtcaggagag agggctccgc ctccgagcgc ctcaaggtcg gggacagcag cagcatcagg 120
cacgagaggg cctccaggag gatgtgcaac ggtggcaggg gccccgccgc caccggcgcg 180
cagtgcgtgc tcacctccga cgccagccca gcagacaccc ttgttctccg gacgtccttc 240
cgcaggaatt acgccgatcc gaacgaggtc gcggccgtca tactcggcgg tggcaccggg 300
actcagctct tccctctcac aagcacaagg gccacacctg ctgttcctat tggaggatgt 360
tacaggctca tcgatattcc catgagcaac tgcttcaata gtggcatcaa caagatattc 420
gtcatgacgc agttcaactc ggcctctctt aatcgtcaca ttcaccgcac ctacctcggc 480
gggggaatca atttcactga tggatctgtt gaggtattgg ccgcgacgca aatgcccggg 540
gaggctgctg gatggttccg cggaacagcg gatgccgtca gaaaatttat ctgggtgctt 600
gaggactatt ataagaataa atccatagag cacattttga tcttgtcggg cgatcagctt 660
tatcgcatgg attacatgga gcttgtgcag aaacatgtgg atgacaatgc tgacattact 720
ttatcatgtg cccctgttgg agagagccgg gcatctgagt acgggctagt gaagttcgac 780
agttcaggcc gtgtggtcca attttctgag aagccaaagg gtgacgatct ggaagcgatg 840
aaagtggaca ccagttttct caatttcgcc atcgacgacc ctgctaaata tccatacatt 900
gcttctatgg gagtctatgt cttcaaaaga gatgttctgc tcaaccttct aaagtcaaga 960
tacgcagaac tacatgactt tgggtctgaa atcctcccga gagctctgca tgaccacaat 1020
gtacaggctt atgtcttcac tgactactgg gaggacattg gaacaatcag atccttcttc 1080
gatgcgaaca tggccctctg cgagcagccc ccaaagttcg agttttatga tcccaaaact 1140
cccttcttca cctcgcctcg atacttgcca ccaacaaagt cagacaagtg caggatcaaa 1200
gaagcgatca tttcgcacgg ctgcttcttg cgtgaatgca aaatcgagca ctccatcatc 1260
ggcgttcgtt cacgcctaaa ctccggaagc gaactcaaga acgcgatgat gatgggtgcg 1320
gattcgtacg agaccgagga tgagatctcg aggctgatgt ccgagggcaa ggtccccatc 1380
ggcgtcgggg agaacacaaa gatcagcaac tgcatcatcg acatgaacgc gaggatagga 1440
agggacgtgg tcatctcaaa caaggaggga gtgcaagaag ccgacaggcc ggaggagggg 1500
tactacatca ggtccgggat cgtggtgatc cagaagaacg cgaccatcaa ggacggcacc 1560
gtcgtgtag 1569
serial number (ID) 2
Length 19
Molecular type DNA
Feature location/qualifier:
- source, 1..19
> mol_type, other DNA
> organism, synthetic construct
residues:
catgtgcaga aagtggaca 19
serial number (ID) 3
Length of 20
Molecular type DNA
Feature location/qualifier:
- source, 1..20
> mol_type, other DNA
> organism, synthetic construct
residues:
agaaggatct gattgttcca 20
serial number (ID) 4
Length 19
Molecular type DNA
Feature location/qualifier:
- source, 1..19
> mol_type, other DNA
> organism, synthetic construct
residues:
tcgctcgctc ttcttcagc 19
serial number (ID) 5
Length of 20
Molecular type DNA
Feature location/qualifier:
- source, 1..20
> mol_type, other DNA
> organism, synthetic construct
residues:
tgccaccgcc gagtatgacg 20
serial number (ID) 6
Length 19
Molecular type DNA
Feature location/qualifier:
- source, 1..19
> mol_type, other DNA
> organism, synthetic construct
residues:
atgcttctca acttctcgt 19
serial number (ID) 7
Length 19
Molecular type DNA
Feature location/qualifier:
- source, 1..19
> mol_type, other DNA
> organism, synthetic construct
residues:
gctgagacga gtgctaacc 19
serial number (ID) 8
Length of 21
Molecular type DNA
Feature location/qualifier:
- source, 1..21
> mol_type, other DNA
> organism, synthetic construct
residues:
catgtggatg acaatgctga c 21
serial number (ID) 9
Length of 21
Molecular type DNA
Feature location/qualifier:
- source, 1..21
> mol_type, other DNA
> organism, synthetic construct
residues:
cttactactg atgctgcctg c 21
serial number (ID) 10
Length 2225
Molecular type DNA
Feature location/qualifier:
- source, 1..2225
> mol_type, other DNA
> organism, synthetic construct
residues:
tcgctcgctc ttcttcagcg cacgtactct acgaggccca cagcccagtc cgtgggcccg 60
ggcgtggcgt gtcgggcagg cggaggccca cagcacagca cagcacagta gtactactgc 120
cacgcccacg ccgcactgta gcacagcaca gcacagctag atgtactacc gctcaaagaa 180
aaaaaaaact aggtgtacta gcacagcggt cgggagctcg ccgagcgatt tccgccgttg 240
gttcccccgg tgatgtgtcg gggcggcagc gcacggggga agggacaaac agacaatggc 300
cgtgtagccg tgtccaggtg agacgaaccg aacgtgcttt gttgccgccg gccggttggg 360
aaaggggatc gatcaattcc gggggccaga gccagcggca cgctgcgcgt ccgtccgtgc 420
gtgcgagacg agccaggcca ggccaggctt tataaccgac ggcaaaaagg aagagcgagc 480
tgggaagccg acggcggcgg cggcgagccg tggccagttg tccttgtcgg atccggtcgt 540
gcgccaacgg agagcattcc acggcaggct ggctgcgggc gtcgagacct gatggcaaca 600
aaagccgcga cggacaccgg aggcgccggc ggtagcgtgg cgggcggaaa ggcgggccgg 660
aggcgtgcgg cgagggccaa gccaagggac ggtgcggggg gcggtaccgt cctgcgcttg 720
ccccgcgcac gcacatggcc gcgggcatgg ggaatggtgg ggggtgggga tagcgccaag 780
agggaaagag agtaacgtag cgactccagt ccgggccatg ccgccctcac gctacgcccc 840
gtgctccctc cctctgctgt ccgggctgct ccacaccgtc cgttctttct cttgcagtgc 900
agtgggaggg acgccaacgc ctccgcccgc tttatatgcc ccgccctcca catcaccaca 960
gcccccgaac tcaacggctc caccgtctct cctctcctct ctctgccctc accaccacca 1020
catcgactag cggggccacc cccggcctct gcctccctgc atttgattca tccgtcgctt 1080
gcccggtcgc ccgtcgatcc gttaacccgc cgtccgtccg cccgtccccg gcagccgcgc 1140
cgcaggtaac aacacatttc cctccctcct tccccccctt gtggattaat ttgcgttccg 1200
gtgatgcgta atggcgttga aaggtggcgg gcgggcgctg cctgttcctc ccgtccggac 1260
cggactccgc cgcgcattaa tgccgcccca ttccggcgcg ggcggggcac gggagcgaca 1320
ggggatcaga cgggtgggcg gttcctcgac ggcaccggcc cgtggggtga gcgagtggca 1380
ccgcgcgccg gccagcggcg tcgttgggca tagccgtggg ccagccgccg gggaggatga 1440
aggagggcgg atcttgttgc cgacttaatt ttaggatcac ggcccctccg tttctggatt 1500
ggaggggccg ggcatgtttg atttatttct cttatctgaa ttaactgacg gcgaccacga 1560
gtaggaggag gatcacggcc ccttccccga ttgttgtatt tggtttgcct tcccgacgac 1620
aaaaatgtga gtcctccttg gatcctgcgc gccctgctca attttcttca agtatttcgg 1680
cgttctcact tctgaccagc tcggtctcat catcactcct ccttaaaata cagtccacat 1740
cttttcccgt gtcaagaagc ttccgtttca ttaacgaaac tctgcactgc actgcaccgc 1800
acttgtgcag tgaaagtctt cttttccact tgcaaagttc atcgggatat tctgcctcgc 1860
ttcctttccc tgcgagttca gaaggggaaa cagcaaagga agaacgccag ccacaagatc 1920
agagggtcat ctcctgattc acgtgaccct gccccgaaaa gtcacaaagt actccacacc 1980
tttcccactc ttccactagt ccttttcccc atgatcattg ttccgaaata ctactactga 2040
gctttacaga gggccgtgac ccgtgaccgt gagagagggg gaaaaagtcc gtgctctttt 2100
tgccccatgt ataaattcat cagtgatttg ggccaagtca tgtgcagtta acggatttct 2160
caaggttcca tggttgctga tgcttctcaa cttctcgtct tgttgcagtt gcaggtggac 2220
tgcga 2225
serial number (ID) 11
Length of 20
Molecular type DNA
Feature location/qualifier:
- source, 1..20
> mol_type, other DNA
> organism, synthetic construct
residues:
taatacgact cactataggg 20
serial number (ID) 12
Length of 24
Molecular type DNA
Feature location/qualifier:
- source, 1..24
> mol_type, other DNA
> organism, synthetic construct
residues:
aagaacatcg attttccatg gcag 24
serial number (ID) 13
Length 3351
Molecular type DNA
Feature location/qualifier:
- source, 1..3351
> mol_type, other DNA
> organism, synthetic construct
residues:
atgtcatcga tgcagttcag cagcgtgctg cccctggagg gcaaggcgtg cgtctccccc 60
gtcaggagag agggctccgc ctgcgagcgc ctcaaggtcg gcgacagcag cagcatcagg 120
cacgacaggg cctccaggag gatgtgcaat ggcggcaggg gccccgccgc caccggcgcg 180
cagtgcgtgc tcacctccga cgccagcccg gcagacaccc ttgtaagttt gatgatcgag 240
ttgcaatgag acactctgtt atgttagcat ctctgatgtt ttcatgcaaa aaaaaataaa 300
aatacctgca ggttctccgg acgtccttcc ggaggaatta cgccgatccg aacgaggtgg 360
cggccgtcat actcggcggt ggcaccggga ctcagctctt cccactcaca agcacaaggg 420
ccacacctgc tgtaagatga cacattcatc atccagttct atgtttttaa ctgtcttgct 480
ctttttctat tacaggttga tagtttaaca ctagcacatc tcagttgcag aaatgaattt 540
tagaaaatct gtgttactta cagatatgtt gtaccgtaga tttcgataaa gggtgcttta 600
ttacttaaag gaagttctac cgtagatgat tcccagtttt agatagatag gaatgaggaa 660
gctttattta aatttcactt gtcatctatc tgcacatggc aattgatttg gataggacca 720
gttgaagtac agctcattca gtagcaagtt ttctgcattg tgctgtgttc aaatcattat 780
tagcagcagt ggtttctgta gccggacccg atgatgcccc ccactttaac cgacatgatg 840
tcaacctcaa ggctatcagt ggccatccat gtagcccgtc cagtcatctt cggatcgcca 900
gcctgaatct tgttcctttc cctttgttgc attgctcagg ttcctattgg aggatgttac 960
aggctcatcg acattcccat gagcaactgc ttcaacagtg gcatcaacaa gatattcgtc 1020
atgacccagt tcaactcggc ctcgcttaat cgccacattc accgcaccta cctcggcggg 1080
ggaatcaatt tcactgatgg atccgttgag gtaattgatc tcagcttatt aatttatact 1140
tgattttttc atcttgacgt cttgggcttt tagctgacta cagcacacat tgactgatac 1200
acctttttgc aacaaatagg tattggccgc gacgcaaatg cccggggagg ctgctggatg 1260
gttccgcgga acagcggacg ccgtcagaaa atttatctgg gtgcttgagg tgagaagagt 1320
actatctgga tacttgtaca tctgcatgtg ttaaagtaat gctacttgtg ttatgtgtac 1380
ctctccagga ctattataag aataaatcca tagagcacat tttgatcttg tcgggcgatc 1440
agctttatcg catggattac atggagcttg tgcaggtttg tgatgttctc tttaaccagc 1500
tccagtgtat gttctgtttt gaactggctg accaagctgt ggcatacatc ctttacagaa 1560
acatgtggat gacaatgctg acattacttt atcatgtgcc cctgttggag agaggtatta 1620
actattgttc tacatcccag atccttattt atgagtgtat cttccttttg ttgaagttgc 1680
actgcatcct gatttgcttt tctttgtttt ggaagccggg catctgagta cgggctagtg 1740
aagttcgaca gttcaggccg tgtggtccag ttttctgaga agccaaaggg tgacgatctg 1800
gaagcgatgg ttagcactcg tctcagctca ctgtgtcaga tatatgccta aagatatgac 1860
tgtcttaaat aagttgcagt atctgaaatt taagccatcg atcatgtgca gaaagtggac 1920
accagttttc tcaatttcgc catagacgat cctgctaaat atccatacat tgcttctatg 1980
ggagtctatg tcttcaaaag agatgttctg ctcaaccttc taaagtaaga acctgctctt 2040
gtggtgacgc cgtcgtcctg tcggtctgaa ttctaatatc cgcattctca ggtcaagata 2100
tgcagaacta catgactttg ggtctgagat cctcccgaga gctctgcatg accacaatgt 2160
acaggtaaac taacagtgtg agcttcattc gttgtttctg ttggggaggt agctctgcgt 2220
gaaatgcagt ctgtaaaact atttcctatg tgttgcacga agtggccgtt catttgacac 2280
tgtttttgtc aggcttatgt cttcactgac tactgggagg acattggaac aatcagatcc 2340
ttcttcgatg caaacatggc cctctgcgag caggtgccat tatctttttg ggtgcttccc 2400
aaatttgact tcagtagtgc taatatgata atcgataaat gataaagtta tattaagact 2460
cttcacttaa tacatttttc agtattttca cacattttgt tcattgcagc ccccaaagtt 2520
cgagttttat gatcccaaaa ctcccttctt cacctcgcct cgatacctgc caccaacaaa 2580
gtcagacaag tgcagggtac gtgttagaat cagcctgcca ttcagactat tccatacaag 2640
cattagctgt aagcatctga tatggattct tctgtgtgca gatcaaagaa gcgatcattt 2700
cgcacggctg cttcttgcgt gaatgcaaaa tcgagcactc catcatcggc gttcgctcac 2760
gcctaaactc cggaagcgag ctcaaggtat gccttttcgc accgaataat ggcatgtggt 2820
gcatgtcaac tgtaaaatca tcctgtgtct gaacgctgtc gctcttccaa atggtggtgg 2880
tttgcagaat gcgatgatga tgggcgcgga ctcgtacgag accgaagacg agatctcgag 2940
gctgatgtcg gagggcaagg tccccatcgg cgtcggggag aacacaaaga tcaggtgaga 3000
gagactgatc tgtacctcag gaacgacacg taccgttcgg cgcaagtgcc acattatctc 3060
ttaacacgtt ttgcgtggat acgttgcagc aactgcatca tcgacatgaa cgcgaggata 3120
ggaagggacg tggtcatctc aaacaaggag gtaagacacc ctccttcaga gttttaaacc 3180
tggtcccaaa gttctgaaga aaacaacact tgtgtctgaa accacgccgc tttgtgccat 3240
ttctgtcagg gagtgcaaga agccgacagg ccggaggagg ggtactacat caggtccggg 3300
atcgtggtga tccagaagaa cgcgaccatc aaggacggca ccgtcgtgta g 3351
serial number (ID) 14
Length 3351
Molecular type DNA
Feature location/qualifier:
- source, 1..3351
> mol_type, other DNA
> organism, synthetic construct
residues:
atgtcatcga tgcagttcag cagcgtgctg cccctggagg gcaaggcgtg cgtctccccc 60
gtcaggagag agggctccgc ctgcgagcgc ctcaaggtcg gcgacagcag cagcatcagg 120
cacgacaggg cctccaggag gatgtgcaat ggcggcaggg gccccgccgc caccggcgcg 180
cagtgcgtgc tcacctccga cgccagcccg gcagacaccc ttgtaagttt gatgatcgag 240
ttgcaatgag acactctgtt atgttagcat ctctgatgtt ttcatgcaaa aaaaaataaa 300
aatacctgca ggttctccgg acgtccttcc ggaggaatta cgccgatccg aacgaggtgg 360
cggccgtcat actcggcggt ggcaccggga ctcagctctt cccactcaca agcacaaggg 420
ccacacctgc tgtaagatga cacattcatc atccagttct atgtttttaa ctgtcttgct 480
ctttttctat tacaggttga tagtttaaca ctagcacatc tcagttgcag aaatgaattt 540
tagaaaatct gtgttactta cagatatgtt gtaccgtaga tttcgataaa gggtgcttta 600
ttacttaaag gaagttctac cgtagatgat tcccagtttt agatagatag gaatgaggaa 660
gctttattta aatttcactt gtcatctatc tgcacatggc aattgatttg gataggacca 720
gttgaagtac agctcattca gtagcaagtt ttctgcattg tgctgtgttc aaatcattat 780
tagcagcagt ggtttctgta gccggacccg atgatgcccc ccactttaac cgacatgatg 840
tcaacctcaa ggctatcagt ggccatccat gtagcccgtc cagtcatctt cggatcgcca 900
gcctgaatct tgttcctttc cctttgttgc attgctcagg ttcctattgg aggatgttac 960
aggctcatcg acattcccat gagcaactgc ttcaacagtg gcatcaacaa gatattcgtc 1020
atgacccagt tcaactcggc ctcgcttaat cgccacattc accgcaccta cctcggcggg 1080
ggaatcaatt tcactgatgg atccgttgag gtaattgatc tcagcttatt aatttatact 1140
tgattttttc atcttgacgt cttgggcttt tagctgacta cagcacacat tgactgatac 1200
acctttttgc aacaaatagg tattggccgc gacgcaaatg cccggggagg ctgctggatg 1260
gttccgcgga acagcggacg ccgtcagaaa atttatctgg gtgcttgagg tgagaagagt 1320
actatctgga tacttgtaca tctgcatgtg ttaaagtaat gctacttgtg ttatgtgtac 1380
ctctccagga ctattataag aataaatcca tagagcacat tttgatcttg tcgggcgatc 1440
agctttatcg catggattac atggagcttg tgcaggtttg tgatgttctc tttaaccagc 1500
tccagtgtat gttctgtttt gaactggctg accaagctgt ggcatacatc ctttacagaa 1560
acatgtggat gacaatgctg acattacttt atcatgtgcc cctgttggag agaggtatta 1620
actattgttc tacatcccag atccttattt atgagtgtat cttccttttg ttgaagttgc 1680
actgcatcct gatttgcttt tctttgtttt ggaagccggg catctgagta cgggctagtg 1740
aagttcgaca gttcaggccg tgtggtccag ttttctgaga agccaaaggg tgacgatctg 1800
gaagcgatgg ttagcactcg tctcagctca ctgtgtcaga tatatgccta aagatatgac 1860
tgtcttaaat aagttgcagt atctgaaatt taagccatcg atcatgtgca gaaagtggac 1920
accagttttc tcaatttcgc catcgacgat cctgctaaat atccatacat tgcttctatg 1980
ggagtctatg tcttcaaaag agatgttctg ctcaaccttc taaagtaaga acctgctctt 2040
gtggtgacgc cgtcgtcctg tcggtctgaa ttctaatatc cgcattctca ggtcaagata 2100
tgcagaacta catgactttg ggtctgagat cctcccgaga gctctgcatg accacaatgt 2160
acaggtaaac taacagtgtg agcttcattc gttgtttctg ttggggagct agctctgcgt 2220
gaaatgcagt ctgtaaaact atttcctatg tgttgcacga agtggccgtt catttgacac 2280
tgtttttgtc aggcttatgt cttcactgac tactgggagg acattggaac aatcagatcc 2340
ttcttcgatg caaacatggc cctctgcgag caggtgccat tatctttttg ggtgcttccc 2400
aaatttgact tcagtagtgc taatatgata atcgataaat gataaagtta tattaagact 2460
cttcacttaa tacatttttc agtattttca cacattttgt tcattgcagc ccccaaagtt 2520
cgagttttat gatcccaaaa ctcccttctt cacctcgcct cgatacttgc caccaacaaa 2580
gtcagacaag tgcagggtac gtgttagaat caacctgcca ttcagactat tccatacaag 2640
cattagctgt aagcatctga tatggattct tctgtgtgca gatcaaagaa gcgatcattt 2700
cgcacggctg cttcttgcgt gaatgcaaaa tcgagcactc catcatcggc gttcgctcac 2760
gcctaaactc cggaagcgag ctcaaggtat gccttttcgc accgaataat ggcatgtggt 2820
gcatgtcaac tgtaaaatca tcctgtgtct gaacgctgtc gctcttccaa atggtggtgg 2880
tttgcagaat gcgatgatga tgggcgcgga ctcgtacgag accgaagacg agatctcgag 2940
gctgatgtcg gagggcaagg tccccatcgg cgtcggggag aacacaaaga tcaggtgaga 3000
gagactgatc tgtacctcag gaacgacacg taccgttcgg cgcaagtgcc acattatctc 3060
ttaacacgtt ttgcgtggat acgttgcagc aactgcatca tcgacatgaa cgcgaggata 3120
ggaagggacg tggtcatctc aaacaaggag gtaagacacc ctccttcaga gttttaaacc 3180
tggtcccaaa gttctgaaga aaacaacact tgtgtctgaa accacgccgc tttgtgccat 3240
ttctgtcagg gagtgcaaga agccgacagg ccggaggagg ggtactacat caggtccggg 3300
atcgtggtga tccagaagaa cgcgaccatc aaggacggca ccgtcgtgta g 3351
END
Claims (7)
- the application of SNP locus in identifying thousand seed weight characters of wheat is characterized in that the SNP locus is positioned at 1944bp of a coding region of a wheat TaAGPL1-1B gene, and polymorphism is A/C;the nucleotide sequence of the coding region of the TaAGPL1-1B gene is shown as SEQ ID No.13 or SEQ ID No. 14.
- 2. The use according to claim 1, wherein the coding region CDS sequence of the TaAGPL1-1B gene is shown in SEQ ID No. 1.
- 3. A method for detecting excellent allelic variation of the TaAGPL1-1B gene in wheat, comprising the steps of:carrying out PCR amplification on genome DNA of the material to be detected by using CAPS molecular marker primer to obtain a PCR amplification product;carrying out enzyme digestion on the PCR amplification product by using restriction enzyme Taq I, and detecting polymorphism of the enzyme digestion product;when the enzyme digestion product comprises two short fragments of 401bp and 42bp, the allelic variation type is TaAGPL1-1B-C; when the enzyme cutting product has no short fragment, the allelic variation type is TaAGPL1-1B-A;the CAPS molecular marker primer pair comprises an upstream primer CAPS-F with a sequence shown as SEQ ID NO.2 and a downstream primer CAPS-R with a sequence shown as SEQ ID NO. 3.
- 4. The method according to claim 3, wherein the reaction system for PCR amplification comprises, in terms of 50. Mu.L: 200ng of genome DNA of a material to be detected, 0.3 mu L of 10 mu M upstream primer CAPS-F, 0.3 mu L of 10 mu M downstream primer CAPS-R, 2X KOD OneTM PCR Master Mix mu L and the balance of ddH 2 O；The PCR amplification reaction is carried out by the following steps: the first stage is pre-denaturation at 94 ℃ for 3min; the second stage of denaturation at 98 ℃ for 10s, annealing at 59 ℃ for 30s, and extension at 68 ℃ for 3min,32 cycles; and a third stage: final extension at 68℃for 5min.
- Application of CAPS molecular marker primer pairs or the method of any one of claims 3-4 in identifying thousand kernel weight types of wheat;the CAPS molecular marker primer pair comprises an upstream primer CAPS-F with a sequence shown as SEQ ID NO.2 and a downstream primer CAPS-R with a sequence shown as SEQ ID NO. 3.
- Application of CAPS molecular marker primer pairs or the method of any one of claims 3-4 in screening high thousand grain weight wheat;the CAPS molecular marker primer pair comprises an upstream primer CAPS-F with a sequence shown as SEQ ID NO.2 and a downstream primer CAPS-R with a sequence shown as SEQ ID NO. 3.
- 7. A method for breeding high thousand grain weight wheat with the assistance of molecular markers is characterized in that CAPS molecular marker primer pairs are utilized to carry out molecular markers on target wheat, the 1944bp position of the coding region sequence of the wheat TaAGPL1-1B gene is screened, and the germplasm with polymorphism C is subjected to subsequent screening;the nucleotide sequence of the coding region of the TaAGPL1-1B gene is shown as SEQ ID No.13 or SEQ ID No. 14;the CAPS molecular marker primer pair comprises an upstream primer CAPS-F with a sequence shown as SEQ ID NO.2 and a downstream primer CAPS-R with a sequence shown as SEQ ID NO. 3.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210269669.5A CN114622031B (en) | 2022-03-18 | 2022-03-18 | SNP locus and CAPS molecular marker primer pair for identifying thousand seed weight characteristics of wheat and application of SNP locus and CAPS molecular marker primer pair |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210269669.5A CN114622031B (en) | 2022-03-18 | 2022-03-18 | SNP locus and CAPS molecular marker primer pair for identifying thousand seed weight characteristics of wheat and application of SNP locus and CAPS molecular marker primer pair |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN114622031A CN114622031A (en) | 2022-06-14 |
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| CN104584901A (en) * | 2015-01-07 | 2015-05-06 | 河南农业大学 | Method for increasing synthesis rate of starch of wheat grain |
| CN105907862A (en) * | 2016-05-05 | 2016-08-31 | 中国科学院遗传与发育生物学研究所 | Method for detecting TaAGPS gene allelic variation relevant to thousand kernel weight |
| CN105950615A (en) * | 2015-12-14 | 2016-09-21 | 中国科学院遗传与发育生物学研究所 | Method and kit for detecting allelic variation of TaAGPL gene |
| CA3049172A1 (en) * | 2016-12-21 | 2018-06-28 | Institute Of Crop Sciences, The Chinese Academy Of Agricultural Sciences | Plant grain trait-related protein, gene, promoter and snps and haplotypes |
| CN111187852A (en) * | 2020-01-16 | 2020-05-22 | 中国科学院遗传与发育生物学研究所 | SNP (single nucleotide polymorphism) site related to wheat yield and application thereof |
| CN111321241A (en) * | 2020-03-07 | 2020-06-23 | 中国科学院遗传与发育生物学研究所农业资源研究中心 | Molecular marker of thousand-grain weight and grain length gene TaGS3-4A of wheat and application thereof |
| CN112176082A (en) * | 2019-07-04 | 2021-01-05 | 中国科学院植物研究所 | SNP molecular marker of wheat grain weight related gene and application thereof |
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| CN104584901A (en) * | 2015-01-07 | 2015-05-06 | 河南农业大学 | Method for increasing synthesis rate of starch of wheat grain |
| CN105950615A (en) * | 2015-12-14 | 2016-09-21 | 中国科学院遗传与发育生物学研究所 | Method and kit for detecting allelic variation of TaAGPL gene |
| CN105907862A (en) * | 2016-05-05 | 2016-08-31 | 中国科学院遗传与发育生物学研究所 | Method for detecting TaAGPS gene allelic variation relevant to thousand kernel weight |
| CA3049172A1 (en) * | 2016-12-21 | 2018-06-28 | Institute Of Crop Sciences, The Chinese Academy Of Agricultural Sciences | Plant grain trait-related protein, gene, promoter and snps and haplotypes |
| CN112176082A (en) * | 2019-07-04 | 2021-01-05 | 中国科学院植物研究所 | SNP molecular marker of wheat grain weight related gene and application thereof |
| CN111187852A (en) * | 2020-01-16 | 2020-05-22 | 中国科学院遗传与发育生物学研究所 | SNP (single nucleotide polymorphism) site related to wheat yield and application thereof |
| CN111321241A (en) * | 2020-03-07 | 2020-06-23 | 中国科学院遗传与发育生物学研究所农业资源研究中心 | Molecular marker of thousand-grain weight and grain length gene TaGS3-4A of wheat and application thereof |
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