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CN-117660675-B - Molecular marker AX-110119937 for detecting zinc content of wheat grains and application thereof

CN117660675BCN 117660675 BCN117660675 BCN 117660675BCN-117660675-B

Abstract

The invention discloses a molecular marker AX-110119937 for detecting zinc content of wheat grains and application thereof, belonging to the field of SNP molecular markers. The AX-110119937 locus is a SNP locus in the wheat genome, such as 36 th nucleotide of SEQ ID No.1, and the nucleotide type is G or A. The invention also provides a method for identifying or assisting in identifying the zinc content character of wheat grains, the method comprises the steps of detecting the genotype of the AX-110119937 locus of the wheat to be detected, wherein the wheat to be detected with the genotype GG of the AX-110119937 locus is zinc-rich wheat, and the zinc content of the grains is higher than or is candidate to be higher than that of the wheat to be detected with the genotype AA of the AX-110119937 locus. The molecular marker provided by the invention can be used for detecting the genotype of QGZnzx.caas-6DL and is used for molecular breeding of zinc-rich wheat.

Inventors

  • HAO YUANFENG
  • SUN MENGJING
  • TONG JINGYANG
  • XIA XIANCHUN
  • HE ZHONGHU

Assignees

  • 中国农业科学院作物科学研究所

Dates

Publication Date
20260505
Application Date
20220907

Claims (8)

  1. 1. The method for identifying or assisting in identifying the zinc content character of the wheat grain is characterized by comprising the steps of detecting the genotype of the wheat to be detected by using a substance for detecting the genotype of an AX-110119937 locus, and identifying or assisting in identifying the zinc content character of the grain of the wheat to be detected according to the genotype of the wheat to be detected; the AX-110119937 locus is a SNP locus in the wheat genome, such as 36 th nucleotide of SEQ ID No.1, the nucleotide type is G or A, The grain zinc content of the wheat to be detected with the genotype GG of the AX-110119937 locus is higher than or the candidate is higher than that of the wheat to be detected with the genotype AA of the AX-110119937 locus; Wherein the genotype is GG and represents the homozygous type of G, the nucleotide type of the AX-110119937 locus in the wheat genome is G, and the genotype is AA and represents the homozygous type of A, the nucleotide type of the AX-110119937 locus in the wheat genome is A.
  2. 2. The method according to claim 1, wherein the substance for detecting genotype at AX-110119937 is A1), A2) or A3): A1 The substance for detecting the genotype of the AX-110119937 locus is a PCR primer composition for amplifying wheat genome DNA fragments including the AX-110119937 locus; a2 The substance for detecting the genotype of the AX-110119937 locus is a PCR reagent containing the PCR primer composition; A3 A kit comprising A1) the PCR primer composition or A2) the PCR reagent.
  3. 3. The method of claim 2, wherein the PCR primer composition comprises a single-stranded DNA having a nucleotide sequence of SEQ ID No.2 at positions 22-40, a single-stranded DNA having a nucleotide sequence of SEQ ID No.3 at positions 22-40, and a single-stranded DNA having a nucleotide sequence of SEQ ID No. 3.
  4. 4. The method according to claim 2 or 3, wherein the PCR primer composition comprises a primer A having a nucleotide sequence shown as SEQ ID No.2, a primer B having a nucleotide sequence shown as SEQ ID No.3 and a primer C having a nucleotide sequence shown as SEQ ID No. 4.
  5. 5. The method of claim 4, wherein the amount of the primer A, primer B and primer C is in a ratio of 2:2:5.
  6. 6. The method according to any one of claims 1 to 5, wherein the method for detecting the genotype of wheat to be tested at the AX-110119937 locus according to claim 1 comprises KASP using the primer composition according to claim 4 as a template to obtain a PCR product, and determining the genotype based on the fluorescent signal of the PCR product.
  7. Use of the AX-110119937 locus or a substance for detecting the genotype of the AX-110119937 locus in any of the following: C1 Identifying or assisting in identifying zinc content traits of wheat kernels; c2 Screening or assisting in screening single wheat plants or lines or varieties with high zinc content in wheat seeds; c3 Auxiliary breeding of zinc content character of wheat seeds; the AX-110119937 locus is a SNP locus in the wheat genome, such as 36 th nucleotide of SEQ ID No.1, the nucleotide type is G or A, The grain zinc content of the wheat to be detected with the genotype GG of the AX-110119937 locus is higher than or the candidate is higher than that of the wheat to be detected with the genotype AA of the AX-110119937 locus; Wherein the genotype is GG and represents the homozygous type of G, the nucleotide type of the AX-110119937 locus in the wheat genome is G, and the genotype is AA and represents the homozygous type of A, the nucleotide type of the AX-110119937 locus in the wheat genome is A.
  8. 8. A method for breeding zinc content traits in wheat grains, which is characterized by selecting wheat with a genotype of GG of AX-110119937 locus as a parent for breeding, wherein the genotype of GG represents homozygosity of G in the nucleotide category of AX-110119937 locus in a wheat genome.

Description

Molecular marker AX-110119937 for detecting zinc content of wheat grains and application thereof Technical Field The invention belongs to the technical field of SNP molecular markers, and particularly relates to a molecular marker AX-110119937 for detecting zinc content of wheat grains and application thereof. Background The lack of trace nutrient elements, also called as 'hidden hunger', especially zinc, causes serious harm to human body, and is a problem which needs to be solved in the global scope. Wheat, one of the world's major food crops, plays an important role in ensuring food safety and nutritional status. However, due to the history selection problem, the zinc content of seeds of modern wheat varieties is generally low, and genetic improvement work of the zinc content of seeds at home and abroad is gradually advancing in order to meet the needs of human health. In recent years, the rapid development of molecular markers provides opportunities for genetic improvement of zinc content in wheat kernels. The molecular marker can be used for effectively identifying genetic loci related to the zinc content of the seeds, and the molecular marker auxiliary selection strategy is used for carrying out genetic improvement on the zinc content of the seeds, so that the selection efficiency is improved, and the breeding period is greatly shortened. Disclosure of Invention The invention aims to solve the technical problem of how to screen or assist in screening zinc-rich wheat. In order to solve the technical problems, the first aspect of the invention provides a method for identifying or assisting in identifying the zinc content traits of wheat seeds, which comprises the steps of detecting the genotype of wheat to be tested by using a substance for detecting the genotype of an AX-110119937 locus, and identifying or assisting in identifying the zinc content traits of the seeds of the wheat to be tested according to the genotype of the wheat to be tested; the AX-110119937 locus is a SNP locus in the wheat genome, such as 36 th nucleotide of SEQ ID No.1, the nucleotide type is G or A, The grain zinc content of the wheat to be detected with the genotype GG of the AX-110119937 locus is higher than or the candidate is higher than that of the wheat to be detected with the genotype AA of the AX-110119937 locus; Wherein the genotype is GG and represents the homozygous type of G, the nucleotide type of the AX-110119937 locus in the wheat genome is G, and the genotype is AA and represents the homozygous type of A, the nucleotide type of the AX-110119937 locus in the wheat genome is A. Further, in the above method, the substance for detecting genotype at AX-110119937 locus is A1), A2) or A3) as follows: A1 The substance for detecting the genotype of the AX-110119937 locus is a PCR primer composition for amplifying wheat genome DNA fragments including the AX-110119937 locus; a2 The substance for detecting the genotype of the AX-110119937 locus is a PCR reagent containing the PCR primer composition; A3 A kit comprising A1) the PCR primer composition or A2) the PCR reagent. Further, in the above method, the PCR primer composition comprises a single-stranded DNA having a nucleotide sequence of SEQ ID No.2 at positions 22-40, a single-stranded DNA having a nucleotide sequence of SEQ ID No.3 at positions 22-40 and a single-stranded DNA having a nucleotide sequence of SEQ ID No. 3. Further, in the above method, the PCR primer composition comprises a primer A having a nucleotide sequence shown as SEQ ID No.2, a primer B having a nucleotide sequence shown as SEQ ID No.3, and a primer C having a nucleotide sequence shown as SEQ ID No. 4. Further, in the above method, the ratio of the amounts of the primer A, primer B and primer C in the PCR primer composition is 2:2:5. Further, the method is characterized by comprising the steps of detecting the genotype of the AX-110119937 locus of the wheat to be tested, and identifying or assisting in identifying the grain zinc content of the wheat according to the genotype, wherein the grain zinc content of the wheat to be tested with the genotype GG of the AX-110119937 locus is higher than or higher than the wheat to be tested with the genotype AA of the AX-110119937 locus. Further, in the method, the method for detecting the genotype of the AX-110119937 locus of the wheat to be detected comprises the steps of taking the genomic DNA of the wheat to be identified as a template, performing KASP by using the PCR primer composition to obtain a PCR product, and determining the genotype according to a fluorescence signal of the PCR product. The method specifically comprises the following operation steps: s1) extracting genome DNA of wheat to be detected; s2) taking the genome DNA extracted in the step S1) as a template, and taking the PCR primer composition as an amplification primer to carry out PCR amplification to obtain a PCR amplification product; S3) judging or assisting in judging the zinc cont