Wheat translocation chromosome as well as molecular marker and application thereof

Abstract

The invention relates to the technical field of crop breeding, and particularly discloses a wheat translocation chromosome and a molecular marker and application thereof, and the translocation chromosome comprises a 1AS. 1AL-1DLDx5 + Dy10 translocation chromosome formed by replacing a 475.8-598.6 Mb segment of a long arm end of a 1A chromosome with a 376.6-498.6 Mb segment of a wheat 1D chromosome long arm end. According to the invention, a 1AS.1AL-1DLDx5 + Dy10 translocation chromosome containing a Dx5 + Dy10 high-quality high-molecular-weight glutenin subunit gene is created, and a matched molecular marker is developed, so that the problems that in existing wheat breeding, excellent genes are lacked, specific translocation line identification depends on a time-consuming and labor-consuming cytological method, and high-throughput application is difficult are solved; according to the method, a 1AS.1 1AL-1DLDx5 + Dy10 translocation chromosome carrier and a non-carrier are rapidly and accurately distinguished, traditional cytological identification is replaced, molecular marker-assisted breeding is supported, and the breeding process of a new variety of high-quality wheat is accelerated.

Description

Technical Field

[0001] This invention belongs to the field of crop breeding technology, specifically relating to a wheat translocation chromosome, its molecular markers, and their applications. Background Technology

[0002] Wheat is the world's most important food crop. With the development of the domestic economy, the improvement of consumption levels, and the adjustment of consumption structure, the genetic improvement of wheat quality is receiving increasing attention. Creating high-quality germplasm resources and cultivating high-quality specialty varieties are of great significance to wheat breeding and production in my country.

[0003] In modern wheat breeding, the lack of superior breeding genes is a major bottleneck restricting further improvement in wheat breeding levels. The gene sources of common wheat are mainly divided into two categories: endogenous genes and exogenous genes from closely related species. Currently, the discovery and utilization of exogenous genes mainly focus on disease resistance and stress tolerance traits, while key breeding genes for complex traits such as yield and quality still primarily originate from endogenous genes. However, regardless of whether endogenous or exogenous genes are utilized, conventional hybridization breeding only changes the combination of genes, not the dosage. Therefore, innovating gene utilization methods is a technical approach to solving the lack of superior genes.

[0004] Common wheat is a typical allochaploid, originating from tetraploid wheat. Triticum turgidum and diploid jointed wheat Aegilops tauschii Hybridization and subsequent spontaneous whole-genome doubling. Changes in chromosome number or structure are common during allopolyploidization. Some chromosomal structural changes, such as chromosomal segment duplication, lead to an increase in the copy number of genes within the duplicated segment. Depending on the gene's sensitivity to dosage, the gene expression level may or may not change with the copy number. Through chromosome engineering, manipulating chromosomal structural variations carrying endogenous genes associated with superior qualities may increase gene expression, thereby leading to alterations in quality-related traits.

[0005] While chromosome engineering can create chromosomal structural variations, theoretically providing a means to increase the dosage of endogenous superior genes, the practical application of this technology faces two major challenges: First, accurately creating specific chromosomal translocation lines containing the target superior gene is a complex and inefficient task; second, there is a lack of molecular markers for rapid, accurate, and high-throughput identification of successfully created specific translocation chromosomes. Currently, the identification of translocation lines heavily relies on time-consuming, labor-intensive, and demanding cytological methods (such as fluorescence in situ hybridization), which significantly limits the efficient screening and application of such superior germplasm resources in large-scale breeding populations.

[0006] Therefore, it is necessary to propose a chromosomal structural variant that can increase the dose of high-quality glutenin subunit genes, and to develop a matching, efficient identification technology that can be used for molecular marker-assisted selection, in order to overcome the above-mentioned defects in existing breeding technologies and accelerate the breeding process of high-quality new wheat varieties. Summary of the Invention

[0007] To address the aforementioned problems, the purpose of this invention is to provide a wheat translocation chromosome, its molecular marker, and its application, through the creation of 1AS.1AL-1DL containing the Dx5+Dy10 high-molecular-weight glutenin subunit gene. Dx5+Dy10 By identifying translocated chromosomes and developing corresponding molecular markers, we can solve the problems of lack of superior genes and reliance on time-consuming and laborious cytological methods for identifying specific translocation lines in existing wheat breeding, which are difficult to apply in high-throughput. We can quickly and accurately distinguish between homozygous, heterozygous, and non-carrier translocations, replace traditional cytological identification, support molecular marker-assisted breeding, and accelerate the breeding process of high-quality new wheat varieties.

[0008] To achieve the above objectives, the technical solution of the present invention is as follows: a wheat translocation chromosome, comprising 1AS.1AL-1DL formed by replacing a segment of the long arm of the wheat 1D chromosome (376.6-498.6 Mb) with a segment of the long arm of the 1A chromosome (475.8-598.6 Mb). Dx5+Dy10 Translocation of chromosomes.

[0009] The principle of the basic scheme is: based on the 1AS.1AL-1DL created in this invention. Dx5+Dy10 The specific structural features of the translocated chromosome (this chromosome is formed by replacing the 475.8-598.6 Mb segment of the long arm of chromosome 1A with the 376.6-498.6 Mb segment of the long arm of chromosome 1D, with recombination breakpoints located in the 475.855823-475.861479 Mb interval of chromosome 1A and the 376.644872-376.821152 Mb interval of chromosome 1D), targeted molecular marker primers were designed to target the specific segment from the recombination breakpoint of chromosome 1A to the centromere, and reverse primers were designed to target the specific segment from the recombination breakpoint of chromosome 1D to the telomere. Only when wheat material carries the specific translocation chromosome can the two primers simultaneously bind to the template sequences on both sides of the recombination breakpoint of the translocation chromosome, and a DNA fragment of 1433 bp in size be specifically amplified through a specific PCR program; materials that do not carry the translocation chromosome cannot amplify the specific band because they lack the complete target sequence for the corresponding primer binding, thus achieving accurate identification and characterization of the target translocation chromosome.

[0010] Furthermore, the location of the translocated chromosome segment is based on IWGSC wheat reference genome version 2.1.

[0011] Furthermore, the 1D chromosome segment contains genes encoding high molecular weight glutenin subunits Dx5 and Dy10.

[0012] Furthermore, 1AS.1AL-1DL Dx5+Dy10 The recombination breakpoint of the translocated chromosome is located at: The 475.855823-475.861479 Mb range on chromosome 1A; and / or the 376.644872-376.821152 Mb range on chromosome 1D.

[0013] Furthermore, 1AS.1AL-1DL Dx5+Dy10 Translocation chromosomes are obtained by utilizing ph1b The chromosome 1A, carrying the high molecular weight glutenin subunit Ax1, was obtained by partial homologous recombination with the chromosome 1D, carrying the high molecular weight glutenin subunits Dx5 and Dy10.

[0014] A molecular marker for wheat translocation chromosomes, comprising the forward primer shown in SEQ ID NO:1 and the reverse primer shown in SEQ ID NO:2, the sequences of which are as follows: Forward primer 1AD-TY-F: 5'-CAGCAGCATGATCGCTGACAC-3'; Reverse primer 1AD-TY-R: 5'-GTTCATGCAGTGTACTTGGAACAG-3'.

[0015] An application of a wheat translocation chromosome in wheat breeding for the selection of new wheat varieties with improved high molecular weight glutenin subunit composition.

[0016] The application of a molecular marker for wheat translocation chromosomes in wheat breeding to assist in the selection of breeding materials carrying the translocation chromosomes.

[0017] Furthermore, the molecular marker is amplified by PCR using the corresponding translocation chromosome as a template: if a specific band of 1433 bp can be amplified, the material is determined to carry the translocation chromosome; if the band cannot be amplified, it is determined not to carry the chromosome.

[0018] Furthermore, the PCR amplification reaction program is as follows: pre-denaturation at 94℃ for 5 minutes; 10 cycles of amplification: denaturation at 94℃ for 30 seconds, annealing at 62-57℃ for 45 seconds (decreasing by 0.5℃ per cycle), extension at 72℃ for 2 minutes; followed by 25 cycles of amplification: denaturation at 94℃ for 30 seconds, annealing at 57℃ for 45 seconds, extension at 72℃ for 2 minutes; and finally, final extension at 72℃ for 7 minutes.

[0019] Compared with the prior art, the present invention can achieve the following technical effects: 1. This molecular marker is based on PCR technology to achieve the targeting of 1AS.1AL-1DL. Dx5+Dy10 The detection of translocation chromosomes is simple to operate and fast, without relying on complex cytological techniques such as fluorescence in situ hybridization. It eliminates the dependence on high-end experimental equipment and professional operating skills, and can quickly complete the screening of large-scale breeding populations, greatly reducing identification time and labor costs, and solving the bottleneck of low efficiency in traditional translocation line identification.

[0020] 2. The forward primer targets the specific region from the 1A chromosome recombination breakpoint to the centromere, and the reverse primer targets the specific region from the 1D chromosome recombination breakpoint to the telomere. Only when the material carries the specific translocation chromosome can the two primers bind simultaneously and amplify a specific 1433 bp band. This product is not produced in non-carrying materials, which can effectively avoid false positive and false negative results. At the same time, it can accurately distinguish between homozygous translocations, heterozygous translocations, and non-carrying materials, providing a reliable basis for the precise selection of target genotypes in breeding.

[0021] 3. This marker can be used to track 1AS.1AL-1DL throughout all generations of wheat breeding. Dx5+Dy10 Translocation and segregation of translocated chromosomes enable targeted screening of materials carrying high-quality Dx5+Dy10 subunit genes, avoiding the loss of desirable traits, significantly improving breeding selection efficiency, and accelerating the breeding process of high-quality new wheat varieties.

[0022] 4. Matching 1AS.1AL-1DL Dx5+Dy10 The translocated chromosome itself carries the Dx5+Dy10 high-molecular-weight glutenin subunit gene, which can increase the dosage of high-quality genes. This molecular marker provides a key tool for its rapid introduction and stable inheritance in breeding, promoting the transformation of this new high-quality germplasm resource into actual breeding results and providing core technical support for the breeding of high-gluten and other high-quality special-purpose wheat varieties. Attached Figure Description

[0023] Figure 1 This invention is 1AS.1AL-1DL Dx5+Dy10 Fluorescence in situ hybridization diagram of translocated chromosomes.

[0024] Figure 2 For the present invention 1AS.1AL-1DL Dx5+Dy10 Materials containing translocated chromosomes (lanes 1 and 3) and materials not containing the 1AS.1AL-1DL of this invention Dx5+Dy10 SDS-PAGE electrophoresis results of translocation chromosome materials (lanes 2 and 4).

[0025] Figure 3 This invention is aimed at 1AS.1AL-1DL Dx5+Dy10Amplification banding results of the specific PCR marker 1AD-TY developed from translocation chromosomes in various materials.

[0026] Figure 4 For the present invention, 1AS.1AL-1DL Dx5+Dy10 Materials homozygous for translocation chromosomes and lacking complete 1A chromosomes were sequenced using a 120k high-performance liquid chromatography-mass spectrometry (HPLC) chip. The sequencing depth changes of SNP sites were analyzed using Python with a 1Mb window.

[0027] Figure 5 This invention is aimed at 1AS.1AL-1DL Dx5+Dy10 The specific PCR marker 1AD-TY developed from translocation chromosomes contains a 1AS.1AL-1DL chromosome. Dx5+Dy10 Amplified banding results of a single translocated chromosome in the F1 population of a cross between a single plant and the common wheat variety Shumai 830. Figure 6 For the present invention, a 1AS.1AL-1DL is included. Dx5+Dy10 Two individual plants (Y4 and Y5) with the target band were amplified in the F1 population of a cross between a translocated chromosome and the common wheat variety Shumai 830, using Urartu wheat ( T. urartu ,2n=2x=14,AA) and jointed wheat ( Ae. tauschii Using genomic DNA (2n=2x=14, DD) as a probe, and utilizing *Aegilops spp.* (a type of goatgrass), a study was conducted. Ae. speltoides 2n=2x=14, SS) represents the results of in situ hybridization of genomes under blocking conditions, where the arrow indicates 1AS.1AL-1DL Dx5+Dy10 chromosome.

[0028] Figure 7 This invention is aimed at 1AS.1AL-1DL Dx5+Dy10 The specific PCR marker 1AD-TY developed from translocation chromosomes contains a 1AS.1AL-1DL chromosome. Dx5+Dy10 The amplified banding results in the self-crossed progeny population of translocated chromosomes.

[0029] Figure 8 For the present invention, a 1AS.1AL-1DL is included. Dx5+Dy10 Cytological identification results of single plants with target bands (A3, A12, and A17) and single plants without target bands (A20, A27, and A31) in the self-crossed progeny population of translocated chromosomes. Detailed Implementation

[0030] The following will describe the implementation of the present invention in detail with reference to the embodiments, so as to fully understand how the present invention uses technical means to solve technical problems and achieve technical effects and to implement it accordingly.

[0031] The specific implementation method is described below with reference to the accompanying drawings.

[0032] Example 1 This embodiment provides a wheat translocation chromosome, comprising 1AS.1AL-1DL formed by replacing a segment of the long arm of the wheat 1D chromosome (376.6-498.6 Mb) with a segment of the long arm (475.8-598.6 Mb) at the end of the long arm of the 1A chromosome. Dx5 +Dy10 Translocation of chromosomes.

[0033] This embodiment also provides a molecular marker for wheat translocation chromosomes, including the forward primer shown in SEQ ID NO:1 and the reverse primer shown in SEQ ID NO:2, the sequences of which are as follows: Forward primer 1AD-TY-F: 5'-CAGCAGCATGATCGCTGACAC-3'; Reverse primer 1AD-TY-R: 5'-GTTCATGCAGTGTACTTGGAACAG-3'.

[0034] The basics are as follows: Figures 1-4 As shown: The experimental process for preparing the above translocation chromosomes and specific molecular markers is as follows: 1. Through ph1b The 1D chromosome, which is induced to express Dx5+Dy10, and the 1A chromosome (carrying the high molecular weight glutenin subunit Ax1) undergo partial homologous chromosome pairing and recombination during meiosis to obtain the 1A-1D translocation selection population. 2. Seeds from the 1A-1D translocation screening population were analyzed using SDS-PAGE to determine the expression of high-molecular-weight glutenin. Materials expressing the Dx1 subunit were discarded. The remaining seeds were germinated and transplanted to the field. Cytological screening was performed to identify individual plants containing the 1AS.1AL-1DL recombinant chromosome. Further self-pollination was conducted, and cytological screening was performed on the progeny to identify plants homozygous for the 1AS.1AL-1DL translocation chromosome and those lacking the complete 1A chromosome. Figure 1 ); 3. DNA was extracted from leaves of the 1AS.1AL-1DL translocation chromosome homozygous material that did not contain the complete 1A chromosome. The DNA was then sequenced using a 120kJ high-performance liquid chromatography (HPLC) chip. Python was used to analyze the sequencing depth changes of SNP sites in a 1Mb window. It was found that the homozygous translocation line lacked the 475.8-598.6 Mb region of the long arm of chromosome 1A, while the sequencing depth of SNP markers in the 376.6-498.6 Mb region of chromosome 1D increased by 1 time. Figure 4Therefore, it is proven that the translocation chromosome was formed by the replacement of chromosome 1A (475.8-598.6 Mb) with chromosome 1D (376.6-498.6 Mb, IWGSC wheat reference genome version 2.1). Figure 4 It was named 1AS.1AL-1DL Dx5+Dy10 .

[0035] 4. Based on the breakpoint interval, design chromosome 1A-specific markers and perform PCR amplification in materials containing and without translocation chromosomes. The presence or absence of bands on agarose gel electrophoresis further identifies the recombination sites of chromosomes 1A and 1D. TraesCS1A03G0702700 Genes (475.858571-475.858599 Mb) and 1D TraesCS1D03G0666700 Between genes (376.656795-376.656823 Mb); 5. Design a specific forward primer 1AD-TY-F located at the recombination site on chromosome 1A to the centromere region and a specific reverse primer 1AD-TY-R located at the recombination site on chromosome 1D to the telomere region to form a specific molecular marker 1AD-TY. Use 1AD-TY to identify carriers and non-carriers of 1AS.1AL-1DL. Dx5+Dy10 PCR identification of wheat with translocated chromosomes showed that only 1AS.1AL-1DL was present. Dx5+Dy10 Wheat with translocated chromosomes could amplify a 1433 bp target-specific band, while wheat materials without translocations did not produce any amplification products. Figure 3 Further investigation was conducted on the carrier 1AS.1AL-1DL. Dx5+Dy10 The amplified products of the translocated chromosome wheat were sequenced and compared with the 1A and 1D reference sequences (IWGSC wheat reference genome version 2.1). Through changes in sequence polymorphism, it was determined that the recombination occurred on chromosome 1A. TraesCS1A03G0702700 and 1D chromosome TraesCS1D03G0666700 The 7th exon of two homologous genes.

[0036] The PCR reaction system and procedure are shown in Table 1 and Table 2.

[0037] Table 1. PCR reaction system

[0038] Table 2. PCR reaction procedure

[0039] Preparation Experiment Results: A novel translocation chromosome (1AS.1AL-1DL) was created, in which the 376.6-498.6 Mb segment of chromosome 1D translocates to the long arm of chromosome 1A, replacing the 475.8-598.6 Mb segment of chromosome 1A. Dx5+Dy10 ). Figure 1 In this context, A represents the use of Urartu wheat ( T. urartu ,2n=2x=14,AA) and jointed wheat ( Ae. tauschii , 2n=2x=14, DD) as probe, using Aegilops spp. ( Ae. speltoides 2n=2x=14, SS) represents the karyotype obtained by blocking in situ hybridization, B represents the fluorescent in situ hybridization karyotype of the translocation-carrying individual plantlet identified by Oligo-pSc119.2 and Oligo-pTa-535, where the arrow indicates 1AS.1AL-1DL Dx5+Dy10 Chromosome analysis and fluorescence in situ hybridization (FISH) showed that the translocated chromosome was formed by the replacement of the end of the long arm of chromosome A with a segment of chromosome D. The Oligo-pSc535 signal characteristics of the chromosome A segment were consistent with those of chromosome 1A, while the Oligo-pSc535 signal characteristics of the chromosome D segment were consistent with those of the end segment of the long arm of chromosome 1D, indicating that it was formed by recombination and exchange at the ends of the long arms of chromosomes 1A and 1D. Figure 2 The 1 and 3 in the text are 1AS.1AL-1DL Dx5+Dy10 Chromosome homozygous material, 2 and 4 are 1AS.1AL-1DL Dx5+Dy10 SDS-PAGE analysis of heterozygous translocation chromosomes and chromosome 1A showed that the homozygous line carrying the translocation chromosome expressed the high-molecular-weight Dx5+Dy10 glutenin subunit, while lacking the Dx1 subunit of the long arm of chromosome 1A, indicating that the 1DL chromosome segment involved in the translocation carried the Dx5+Dy10 subunit.

[0040] Figure 3 Lanes 1-2 and 5-6: Carry 1AS.1AL-1DL Dx5+Dy10 Translocation chromosome material, lanes 3-4: excluding 1AS.1AL-1DL Dx5+Dy10 Homozygous material of chromosome 1A from translocation chromosomes, lane 7: standard DNA (marker).

[0041] Figure 4Figure A shows the sequencing depth changes of the SNP sites on chromosome 1A, and Figure B shows the sequencing depth changes of the SNP sites on chromosome 1D. High-performance liquid chromatography (HPLC) analysis of wheat at 120kJ / m² showed that the homozygous lines carrying this translocation chromosome lacked molecular markers in the 475.8-598.6 Mb region of chromosome 1A, while the concentration in the 376.6-498.6 Mb region of chromosome 1D increased by a factor of 1. The precise structure of this translocation chromosome was confirmed by fluorescence in situ hybridization (FISH) and molecular identification.

[0042] By combining microarray sequencing results and the location of translocation chromosome breakpoints, a novel 1AS.1AL-1DL assay was developed specifically for this invention. Dx5+Dy10 The specific molecular marker 1AD-TY for translocation chromosomes can exclusively identify whether wheat materials carry the 1AS.1AL-1DL invented in this invention. Dx5+Dy10 Translocation chromosomes, enhancing 1AS.1AL-1DL Dx5+Dy10 Breeding screening efficiency of translocation chromosomes and their use for carrying 1AS.1AL-1DL Dx5+Dy10 Identification of translocation materials.

[0043] Experimental Example 1 The basics are as follows: Figure 5 and Figure 6 As shown: For 1AS.1AL-1DL Dx5+Dy10 Experiments to validate translocation chromosome-specific molecular markers in translocation segregating populations.

[0044] The experimental procedure is as follows: 1. Molecular cytological identification revealed a normal 1A and 1AS.1AL-1DL. Dx5+Dy10 The hybrid single plant was crossed with the common wheat variety Shumai 830 to obtain hybrid F1; 2. The seven hybrid seeds obtained were processed using 1AS.1AL-1DL. Dx5+Dy10 The translocation chromosome-specific molecular marker 1AD-TY was amplified by PCR and subjected to agarose gel electrophoresis. For example... Figure 5 As shown, two of the individual plants (Y4 and Y5) amplified a specific band of 1433 bp, while the remaining individual plants did not produce any amplification products. 3. For the two single plants (Y4 and Y5) with target bands amplified by electrophoresis, Urartu wheat ( T. urartu ,2n=2x=14,AA) and jointed wheat ( Ae. tauschii Using genomic DNA (2n=2x=14, DD) as a probe, and utilizing *Aegilops spp.* (a type of goatgrass), a study was conducted. Ae. speltoides 2n=2x=14, SS) was used as a blocking agent for in situ hybridization of the genome. The results showed that both Y4 and Y5 contained a 1AS.1AL-1DL line. Dx5+Dy10 Translocation chromosomes ( Figure 6(This further illustrates the design of the 1AS.1AL-1DL of the present invention.) Dx5+Dy10 Primers for translocation chromosome-specific molecular markers are accurate, specific, and stable.

[0045] Experimental Example 2 The difference from the above experimental examples is that, for example, Figure 7 and Figure 8 As shown: Validation of 1AS.1AL-1DL in different backgrounds Dx5 +Dy10 Experiments on the specificity and stability of translocation chromosome breakpoint markers.

[0046] The experimental procedure is as follows: Step 1, Molecular cytological identification to obtain a normal 1A and 1AS.1AL-1DL. Dx5+Dy10 A heterozygous single plant, which self-pollinates to obtain offspring seeds; Step 2: Randomly select 36 seeds and use 1AS.1AL-1DL Dx5+Dy10 The translocation chromosome-specific molecular marker 1AD-TY was amplified by PCR and subjected to agarose gel electrophoresis. For example... Figure 7 As shown, 30 of the individual strains amplified a specific band of 1433 bp, while the remaining individual strains did not produce any amplification products. Step 3: Randomly select 3 individual plants with target bands (A3, A12, and A17) and 3 individual plants without target bands (A20, A27, and A31) using Urartu wheat ( T. urartu ,2n=2x=14,AA) and jointed wheat ( Ae. tauschii , 2n=2x=14, DD) as probe, using Aegilops spp. ( Ae. speltoides Genomic in situ hybridization was performed using 2n=2x=14, SS) as the blocking method. Simultaneously, fluorescence in situ hybridization was performed using Oligo-pSc119.2 and Oligo-pSc535 probes for identification. Results are shown in […]. Figure 8 The arrow indicates 1AS.1AL-1DL Dx5+Dy10 chromosome.

[0047] A3, A12, and A17 contain 1AS.1AL-1DL Dx5+Dy10 Translocated chromosomes A20, A27, and A31 do not contain 1AS.1AL-1DL. Dx5+Dy10 Translocation chromosomes ( Figure 8 This describes the design of the 1AS.1AL-1DL of the present invention. Dx5+Dy10 The molecular marker primers for translocation chromosome breakpoints are accurate and stable.

[0048] The foregoing description illustrates and describes several preferred embodiments of the invention. However, as previously stated, it should be understood that the invention is not limited to the forms disclosed herein and should not be construed as excluding other embodiments. It can be used in various other combinations, modifications, and environments, and can be altered within the scope of the inventive concept described herein through the foregoing teachings or techniques or knowledge in related fields. Any modifications and variations made by those skilled in the art that do not depart from the spirit and scope of the invention should be within the protection scope of the appended claims.

Claims

1. A wheat translocation chromosome, characterized in that, 1AS.1AL-1DL including a segment of the long arm of chromosome 1A replaced by a segment of the long arm of chromosome 1D from 475.8-598.6 Mb Dx5+Dy10 Translocation chromosome.

2. The translocation chromosome of claim 1, wherein, The segment position of the translocation chromosome is based on the IWGSC wheat reference genome version 2.

1.

3. The translocation chromosome of claim 2, wherein, The 1D chromosome segment comprises genes encoding high-molecular-weight glutenin subunits Dx5 and Dy10.

4. The translocation chromosome of claim 3, wherein, 1 AS.1 AL-1 DL Dx5+Dy10 The recombinant breakpoints of the translocation chromosomes are located at: the 475.855823-475.861479 Mb interval of chromosome 1A; and / or the 376.644872-376.821152 Mb interval of chromosome 1D.

5. The translocation chromosome of claim 4, wherein, 1 AS. 1 AL-1 DL Dx5+Dy10 Translocation chromosomes were obtained by using ph1b The 1A chromosome carrying the high molecular weight glutenin subunit Ax1 was induced to undergo partial homologous recombination with the 1D chromosome carrying the high molecular weight glutenin subunits Dx5 and Dy10 by the gene.

6. A molecular marker of a wheat translocation chromosome, characterized in that, The molecular marker comprises a forward primer shown in SEQ ID NO: 1 and a reverse primer shown in SEQ ID NO: 2, and the sequences of the forward primer and the reverse primer are as follows: Forward primer 1AD-TY-F: 5'-CAGCAGCATGATCGCTGACAC-3'; Reverse primer 1AD-TY-R: 5'-GTTCATGCAGTGTACTTGGAACAG-3'.

7. The translocation chromosome of any one of claims 1-5 for use in wheat breeding, for breeding new wheat varieties with improved high-molecular-weight glutenin subunit composition.

8. The molecular marker of claim 6 for use in wheat breeding, for assisting selection of breeding materials carrying the translocation chromosome.

9. Use of a molecular marker according to claim 8 in wheat breeding, characterized in that, PCR amplification of the genomic DNA of the material to be detected is performed using the molecular marker: if a specific band of 1433 bp can be amplified, it is determined that the material carries the translocation chromosome; if the band cannot be amplified, it is determined that it does not carry the translocation chromosome.

10. Use of a molecular marker according to claim 9 in wheat breeding, characterized in that, The reaction procedure for the PCR amplification is as follows: pre-denaturation at 94℃ for 5 minutes; amplification for 10 cycles: denaturation at 94℃ for 30 seconds, annealing at 62-57℃ for 45 seconds (reducing by 0.5℃ for each cycle), extension at 72℃ for 2 minutes; followed by amplification for 25 cycles: denaturation at 94℃ for 30 seconds, annealing at 57℃ for 45 seconds, extension at 72℃ for 2 minutes; finally, final extension at 72℃ for 7 minutes.

Images