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CN-122012768-A - SNP marker of stem basal rot resistant site Qcr.hnk-6BS of wheat Cheng Zhu stage, KASP detection primer and application

CN122012768ACN 122012768 ACN122012768 ACN 122012768ACN-122012768-A

Abstract

The invention belongs to the technical field of biology, and particularly relates to an SNP marker of a stem rot resistance locus Qcr.hnk-6BS of wheat Cheng Zhu stage, a KASP detection primer and application thereof. The invention provides a product for detecting SNP AX-109329176 at wheat 6BS chromosome 4885850 bp, which comprises (1) a KASP detection primer for detecting SNP AX-109329176 genotype and (2) a reagent or a kit containing the detection primer in (1). The invention also provides application of the product in any aspect of (1) auxiliary breeding of wheat with stem rot resistance in the adult stage, and particularly can be used for auxiliary selection of wheat stem rot resistance breeding offspring materials, and (2) identification or evaluation of stem rot resistance in the adult stage of wheat germplasm resources.

Inventors

  • DU XIJUN
  • XU WEIGANG
  • DING CHAOJIE
  • ZHANG WANWAN
  • QI XUELI
  • HU LIN
  • FANG YUHUI

Assignees

  • 河南省作物分子育种研究院

Dates

Publication Date
20260512
Application Date
20260112

Claims (8)

  1. The use of SNP AX-109329176 in any one of the following aspects, characterized in that, (1) Auxiliary breeding of stem rot resistant wheat in adult stage; (2) Identifying or evaluating the resistance of wheat germplasm resources to stem basal rot in the adult stage; The SNP AX-109329176 is located at 4885850 bp on wheat chromosome 6BS, where the nucleotide polymorphism is T or C.
  2. 2. A product for detecting SNP AX-109329176, characterized in that the SNP AX-109329176 is located at 4885850 bp on wheat chromosome 6BS, where the nucleotide polymorphism is T or C; The product comprises: (1) A KASP detection primer for detecting the SNP AX-109329176 genotype, the KASP detection primer comprising: the upstream primer CF1:5'-AGCGCTCCATTGTTGGGTGTT-3'; The upstream primer CF2:5'-AGCGCTCCATTGTTGGGTGTC-3'; A downstream primer CR 5'-AACGCTAGCCCAGTCGAATAC-3'; (2) A reagent or kit comprising the detection primer described in (1).
  3. 3. The product for detecting SNP AX-109329176 according to claim 2, wherein the 5 'end of the upstream primer CF1 is further provided with a specific fluorescent sequence FAM and the 5' end of the upstream primer CF2 is further provided with a specific fluorescent sequence HEX.
  4. 4. The product for detecting SNP AX-109329176 according to claim 3, wherein, The specific fluorescent sequence FAM is 5'-GAAGGTGACCAAGTTCATGCT-3'; The specific fluorescent sequence HEX is 5'-GAAGGTCGGAGTCAACGGATT-3'.
  5. 5. The use of a product for detecting SNP AX-109329176 according to any one of claims 2 to 4, wherein, (1) Auxiliary breeding of stem rot resistant wheat in adult stage; (2) And (5) identifying or evaluating the resistance of the wheat germplasm resources to the stem basal rot in the adult stage.
  6. 6. A method for detecting the resistance of wheat Cheng Zhu to stem basal rot, comprising: detecting genotype of SNP AX-109329176 in wheat genome, determining resistance of wheat to Cheng Zhu stage stem basal rot according to genotype, wherein SNP AX-109329176 is located at 4885850 bp on wheat chromosome 6BS, polymorphism of SNP marker is T or C, when genotype of SNP AX-109329176 is TT, it is wheat for adult stage stem basal rot, and when genotype of SNP AX-109329176 is CC, it is wheat for adult stage stem basal rot resistance.
  7. 7. The method for detecting the resistance to the stem basal rot of wheat Cheng Zhu according to claim 6, The product for detecting SNP AX-109329176 according to claim 3 or 4 is used for PCR amplification of genomic DNA of wheat, and fluorescence signal detection of PCR amplified product is carried out, red fluorescence indicates CC disease resistance genotype, blue fluorescence indicates TT disease genotype, and green fluorescence indicates CT heterozygous genotype.
  8. 8. A method for breeding wheat, comprising: (1) Detecting the resistance of wheat parent adult stage stem basal rot by the method of claim 6 or 7; (2) And selecting corresponding wheat parents according to breeding targets for breeding.

Description

SNP marker of stem basal rot resistant site Qcr.hnk-6BS of wheat Cheng Zhu stage, KASP detection primer and application Technical Field The invention belongs to the technical field of biology, and particularly relates to an SNP marker of a wheat Cheng Zhu-stage stem basal rot resistance locus Qcr.hnk-6BS, a KASP detection primer and application thereof. Background Wheat (Triticum aestivum l.) is one of the staple food grains for humans. The stem-based rot has the characteristics of strong pathogen rot, wide host, main disease parts of the stem base, disease occurrence in the whole growth period and the like, has extremely great harm to wheat, is difficult to effectively and radically cure by adopting traditional methods such as bactericides and the like, and is the most effective and environment-friendly measure for preventing and controlling the occurrence of the stem-based rot of the wheat. The excavation of the resistance gene and the development of the molecular marker are the basis and the guarantee of wheat stem basal rot resistance breeding. At present, the stem rot resistant wheat breeding creates favorable disease conditions through a field artificial inoculation mode, the case of offspring selection according to phenotype identification is few, and the auxiliary selection breeding is carried out through molecular marker detection. The molecular marker assisted selection is one of the strategies of modern plant breeding, the types and the numbers of the wheat stem rot resistance genes and the QTLs of offspring individuals can be detected and selected from the molecular level in early generations, and the single plants for polymerizing a plurality of target genes and the QTLs can be selected, so that the effectiveness and the durability of the stem rot resistance can be improved. With the rapid development of genome sequencing technology, tens of molecular markers have been generated for application in various scientific researches. Single Nucleotide Polymorphism (SNP) markers are favored in various research fields because of their numerous, widely distributed, dense, and suitable for rapid, high-throughput genotyping. While the research of wheat genomics is rapidly developed, the iterative upgrading of the gene chip technology is promoted, wheat SNP chips with different densities such as 9K, 15K, 55K, 90K, 660K, 820K and the like are developed successively, and based on SNP loci, the development of KASP markers which facilitate the high-throughput detection of the SNP loci is the mainstream of molecular marker development in a considerable period of time at present and in the future. Disclosure of Invention The invention provides an SNP marker SNP AX-109329176 related to the resistance of wheat Cheng Zhu stage stem basal rot, and a KASP detection primer for detecting the SNP marker. The SNP marker and the KASP detection primer can be used for screening of germplasm resources of wheat for resisting stem rot and selecting breeding offspring materials. The invention adopts the following technical scheme: the present invention first provides the use of SNP AX-109329176 in any of the following aspects: (1) The method can be used for auxiliary breeding of wheat with stem rot resistance in the adult period, and can be particularly used for auxiliary selection of the wheat stem rot resistance breeding offspring materials. (2) And (5) identifying or evaluating the resistance of the wheat germplasm resources to the stem basal rot in the adult stage. The SNP AX-109329176 is located at 4885850 bp on wheat chromosome 6BS, where the nucleotide polymorphism is T or C. The present invention provides a product for detecting the above SNP AX-109329176, comprising: (1) A KASP detection primer for detecting the genotype of the above SNP AX-109329176, the KASP detection primer comprising: An upstream primer CF1:5'-AGCGCTCCATTGTTGGGTGTT-3' (SEQ ID NO: 6); an upstream primer CF2:5'-AGCGCTCCATTGTTGGGTGTC-3' (SEQ ID NO: 7); The downstream primer CR 5'-AACGCTAGCCCAGTCGAATAC-3' (SEQ ID NO: 3). The 5 'end of the upstream primer CF1 is also provided with a specific fluorescent sequence FAM, and the 5' end of the upstream primer CF2 is also provided with a specific fluorescent sequence HEX. Wherein, the specific fluorescent sequence FAM is 5'-GAAGGTGACCAAGTTCATGCT-3' (SEQ ID NO: 4); The specific fluorescent sequence HEX is 5'-GAAGGTCGGAGTCAACGGATT-3' (SEQ ID NO: 5). (2) A reagent or kit comprising the primer combination of (1). The invention provides the use of a product for detecting the above SNP AX-109329176 in any of the following aspects: (1) Auxiliary breeding of stem rot resistant wheat in adult stage; (2) And (5) identifying or evaluating the resistance of the wheat germplasm resources to the stem basal rot in the adult stage. The invention provides a method for detecting the resistance of wheat Cheng Zhu-stage stem basal rot, which comprises the following steps: detecting the genotype of the SNP AX-109329176 in the gen