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CN-117660676-B - Molecular marker for detecting zinc content, iron content and thousand kernel weight-cause multiple-effect sites of wheat kernels and application thereof

CN117660676BCN 117660676 BCN117660676 BCN 117660676BCN-117660676-B

Abstract

The invention discloses a molecular marker for detecting a multi-effect site of zinc content, iron content and thousand grain weight of wheat grains and application thereof, and belongs to the technical field of biology. The invention aims to solve the technical problem of identifying or assisting in identifying the thousand grain weight/zinc content/iron content characteristics of wheat grains. In order to solve the technical problems, the invention provides an application of an AX-109500250 locus and a substance for detecting the genotype of the AX-109500250 locus in identifying or assisting in identifying thousand grain weight/zinc content/iron content characters of wheat grains, wherein the AX-109500250 locus is an SNP locus in a wheat genome, such as 48 th nucleotide of SEQ ID No.1, and the nucleotide type is C or T. The molecular marker provided by the invention has important application value for high-zinc/high-iron/high-grain-weight wheat molecular breeding.

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 (7)

  1. Use of an AX-109500250SNP locus and a substance that detects the genotype of an AX-109500250SNP locus in any of the following: A1 Identification or auxiliary identification of thousand kernel weight, zinc content and/or iron content of wheat kernels; a2 Screening or assisting in screening individual or strain or variety of wheat with high thousand kernel weight, zinc content and/or iron content; A3 Wheat kernel thousand kernel weight, zinc content and/or iron content to assist in breeding; the AX-109500250SNP locus is one SNP locus in the wheat genome, is the 48 th nucleotide of SEQ ID No.1, and has the nucleotide type of C or T.
  2. 2. The method according to claim 1, wherein the substance for detecting polymorphism or genotype of the SNP site of AX-109500250 is any one of the following: b1 The substance for detecting the genotype of the AX-109500250SNP locus is a PCR primer composition for amplifying wheat genome DNA fragments including the AX-109500250SNP locus; b2 The material for detecting the genotype of the SNP locus of AX-109500250 is a PCR reagent containing the PCR primer composition of B1); b3 A kit comprising B1) the PCR primer composition or B2) the PCR reagent.
  3. 3. The method according to claim 2, wherein the PCR primer composition comprises a single-stranded DNA having a nucleotide sequence of SEQ ID No.2 at positions 22-41, a single-stranded DNA having a nucleotide sequence of SEQ ID No.3 at positions 22-41, and a single-stranded DNA having a nucleotide sequence of SEQ ID No. 4.
  4. 4. The method according to claim 2 or 3, wherein the PCR primer composition comprises a primer L1 having a nucleotide sequence shown in SEQ ID No.2, a primer L2 having a nucleotide sequence shown in SEQ ID No.3, and a primer R having a nucleotide sequence shown in SEQ ID No. 4.
  5. 5. A method for identifying or assisting in identifying the thousand kernel weight, zinc content and/or iron content traits of wheat seeds, which comprises detecting the genotype of the AX-109500250SNP locus in claim 1 of the wheat to be tested, identifying or assisting in identifying the thousand kernel weight, zinc content and/or iron content of the wheat according to the genotype, wherein the genotype of the AX-109500250SNP locus is the wheat to be tested with the thousand kernel weight, zinc content and/or iron content of TT higher than or candidate higher than the genotype of the AX-109500250SNP locus as the genotype of CC, wherein the genotype is the homozygous type with the TT indicating the nucleotide type of T of the AX-109500250SNP locus in the wheat genome, and the genotype is the homozygous type with the CC indicating the nucleotide type of C of the AX-109500250SNP locus in the wheat genome.
  6. 6. The method according to claim 5, wherein the method for detecting the genotype of the AX-109500250SNP locus in claim 1 of wheat to be tested comprises KASP using the PCR primer composition of claim 4 as a template to obtain a PCR product, and determining the genotype based on the fluorescent signal of the PCR product.
  7. 7. A method for breeding thousand grain weight, zinc content and/or iron content characteristics of wheat seeds is characterized by comprising the step of selecting wheat with genotype TT of an AX-109500250SNP locus as defined in claim 1 as a parent for breeding, wherein the genotype TT represents homozygosity of the nucleotide type T of the AX-109500250 locus in a wheat genome.

Description

Molecular marker for detecting zinc content, iron content and thousand kernel weight-cause multiple-effect sites of wheat kernels and application thereof Technical Field The invention relates to the technical field of biology, in particular to a molecular marker for detecting multiple effect sites of zinc content, iron content and thousand kernel weight of wheat kernels and application thereof. Background Micronutrient deficiency, also known as "implicit starvation," has become the most common public health problem worldwide. Wheat, one of the most widely planted crops in the world, provides about 20% of calories and proteins for humans. Wheat is the main source of daily zinc and iron intake for poor areas relying on grain as the principal food. However, the zinc and iron content of seeds of most of the promoted wheat varieties in the world cannot meet the target requirement, which is probably due to the fact that the past breeding is biased in yield, the nutrition quality is not paid attention, and a large number of excellent genes are lost in the breeding process. Therefore, the genetic locus and important genes of zinc and iron are discovered, and the genetic locus and important genes are introduced into modern wheat varieties, so that the genetic locus and the important genes have important significance for improving the zinc and iron content of wheat seeds and further improving the condition of trace nutrient deficiency. Some studies have shown that there is a "dilution effect" between grain zinc iron concentration and grain yield. While development of trace element rich varieties is an important goal of a wheat breeding program, the sacrifice of grain yield should not be traded for concentration of trace elements. Therefore, it is important to consider both yield and micronutrients in breeding. The molecular marker assisted selective breeding can pyramid a plurality of favorable alleles to achieve the aim of improving a plurality of characters at the same time, but the process is more complicated. Conversely, it may be easier to select for multiple traits simultaneously due to multiple pleiotropic sites. Therefore, the development of molecular markers of multiple-effect sites of zinc content, iron content and thousand grain weight of the grain has important application value for improving zinc-iron content and grain weight of the grain at the same time. Disclosure of Invention The invention aims to solve the technical problem of identifying or assisting in identifying the thousand grain weight/zinc content/iron content characteristics of wheat grains. To solve the above technical problem, in a first aspect, the present invention provides the use of a substance for detecting genotype of AX-109500250 locus in any one of the following: a1 Identifying or assisting in identifying the thousand kernel weight, zinc content and/or iron content traits of the wheat kernel; a2 Screening or assisting in screening individual or strain or variety of wheat with high thousand kernel weight, zinc content and/or iron content; A3 Wheat assisted breeding; The AX-109500250 locus is a SNP locus in the wheat genome, such as 48 th nucleotide of SEQ ID No.1, and the nucleotide type is C or T. In the application, the grain thousand weight, zinc content and/or iron content of the wheat to be detected with the genotype of the AX-109500250 locus being TT are higher than or candidate to be higher than those of the wheat to be detected with the genotype of the AX-109500250 locus being CC, wherein the genotype is a homozygous type with the nucleotide type of the AX-109500250 locus being T in a wheat genome, and the genotype is a homozygous type with the nucleotide type of the AX-109500250 locus being C in the wheat genome. Further, in the above application, the substance for detecting polymorphism or genotype of AX-109500250 site is any one of the following: B1 The substance for detecting genotype of AX-109500250 locus contains PCR primer composition for amplifying wheat genome DNA fragment including AX-109500250 locus; B2 The substance for detecting the genotype of the AX-109500250 locus is a PCR reagent containing the PCR primer composition; b3 A kit comprising B1) the PCR primer composition or B2) the PCR reagent. Further, in the above application, the PCR primer composition comprises a single-stranded DNA having a nucleotide sequence of SEQ ID No.2 at positions 22-41, a single-stranded DNA having a nucleotide sequence of SEQ ID No.3 at positions 22-41 and a single-stranded DNA having a nucleotide sequence of SEQ ID No. 4. Further, in the above application, the PCR primer composition comprises a primer L1 with a nucleotide sequence shown as SEQ ID No.2, a primer L2 with a nucleotide sequence shown as SEQ ID No.3, and a primer R with a nucleotide sequence shown as SEQ ID No. 4. Further, the 5' ends of the primer L1 and the primer L2 are specific fluorescent tag sequences. Further, the specific fluorescent tag sequences of the 5' en