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CN-118240967-B - Molecular marker and method for identifying drought resistance of wheat in seedling stage

CN118240967BCN 118240967 BCN118240967 BCN 118240967BCN-118240967-B

Abstract

The invention discloses a molecular marker and a method for identifying drought resistance of wheat in seedling stage. The molecular marker for identifying drought resistance of wheat in seedling stage is a 1038 th nucleotide corresponding to SEQ ID No.1 in a sequence table in wheat genome, and is A or G. Experiments prove that the molecular marker is related to the drought resistance of wheat, and the drought resistance of homozygous wheat with the nucleotide A corresponding to the 1038 th position of SEQ ID No.1 in the genome is greater than that of homozygous wheat with the nucleotide G corresponding to the 1038 th position of SEQ ID No.1 in the genome. The molecular marker can be used for rapidly and accurately finding out wheat with high drought resistance in the seedling stage, provides a novel method for wheat molecular marker assisted selection breeding, and has important significance in culturing drought-resistant high-yield wheat varieties or research.

Inventors

  • LI CHAONAN
  • JING RUILIAN
  • Fan Zipei
  • MAO XINGUO
  • WANG JINGYI
  • LI LONG

Assignees

  • 中国农业科学院作物科学研究所

Dates

Publication Date
20260505
Application Date
20240509

Claims (8)

  1. 1. The application of a substance for detecting wheat molecular markers in detecting or assisting in detecting drought resistance of wheat is characterized in that the wheat molecular markers are nucleotides corresponding to 1038 th position of SEQ ID No.1 in a sequence table in a wheat genome, and are A or G.
  2. 2. The method according to claim 1, wherein the substance for detecting the molecular markers of wheat comprises a Primer pair named TaNRT2-Primer-BsmaI or a Primer pair named TaNRT 2-6A-Primer; the TaNRT-Primer-BsMI consists of two single-stranded DNAs shown as SEQ ID No.2 and SEQ ID No.3 in a sequence table; The TaNRT-6A-Primer consists of two single-stranded DNAs shown as SEQ ID No.4 and SEQ ID No.5 in a sequence table.
  3. 3. The method of claim 2, wherein the substance further comprises the restriction enzyme BsmAI.
  4. 4. The method according to claim 1 to 3, wherein the drought resistance of the wheat whose nucleotide A corresponding to the 1038 th position of SEQ ID No.1 in the sequence Listing is greater than or alternatively greater than the drought resistance of the wheat whose nucleotide G corresponding to the 1038 th position of SEQ ID No.1 in the genome.
  5. 5. A method for detecting or assisting in detecting drought resistance of wheat, comprising detecting the wheat molecular marker in claim 1, and determining the drought resistance of wheat according to the method that the drought resistance of wheat with the nucleotide A corresponding to 1038 position of SEQ ID No.1 in a genome is greater than or is candidate to be greater than the drought resistance of wheat with the nucleotide G corresponding to 1038 position of SEQ ID No.1 in the genome.
  6. 6. The method according to claim 5, wherein the detection of the wheat molecular marker according to claim 1 is performed using the substance for detecting a wheat molecular marker according to any one of claims 1 to 3.
  7. 7. Any of the following applications: use of the wheat molecular marker of X1) claim 1 in wheat drought resistance breeding; x2) the application of the wheat molecular marker in the method in the detection or auxiliary detection of drought resistance of wheat; Use of X3) a substance for detecting a wheat molecular marker according to any one of claims 1-3 in wheat drought resistance breeding; Use of X4) a substance for detecting a wheat molecular marker according to any one of claims 1 to 3 for the preparation of a wheat drought resistance breeding product; use of X5) a substance for detecting a wheat molecular marker according to any one of claims 1-3 for the preparation of a product for detecting or aiding in the detection of drought resistance in wheat; X6) use of the method of claim 5 or 6 in wheat drought resistance breeding; X7) detecting the substances corresponding to 1038 nucleotide of SEQ ID No.1 in a sequence table in a wheat genome, and the application of the substances in breeding wheat with excellent drought resistance; x8) detecting substances corresponding to 1038 th nucleotide of SEQ ID No.1 in a sequence table in a wheat genome, and preparing and breeding products of wheat with excellent drought resistance.
  8. 8. The wheat drought resistance breeding method includes detecting 1038 th nucleotide corresponding to SEQ ID No.1 in the sequence list in wheat genome and selecting wheat with 1038 th nucleotide corresponding to SEQ ID No.1 in the sequence list as parent for breeding.

Description

Molecular marker and method for identifying drought resistance of wheat in seedling stage Technical Field The invention relates to a molecular marker and a method for identifying drought resistance of wheat in seedling stage in the field of biotechnology. Background Wheat is one of three main grain crops in China, and high and stable yield of the wheat is important to guaranteeing national grain safety. The drought and water shortage seriously affects the wheat production, and the improvement of the drought resistance of the wheat is an important means for ensuring high and stable yield. Along with the development of molecular biology, molecular marker assisted breeding provides a convenient and quick method for selecting target characters, explores excellent gene resources and develops molecular markers, and lays a foundation for efficient genetic improvement and germplasm innovation. Disclosure of Invention The invention aims to solve the technical problem of how to detect the drought resistance of wheat. In order to solve the technical problems, the invention firstly provides application of a substance for detecting wheat molecular markers in detecting or assisting in detecting drought resistance of wheat; The wheat molecular marker is a nucleotide corresponding to 1038 position of SEQ ID No.1 in a sequence table in a wheat genome, and is A or G. In the above application, the substance for detecting the wheat molecular marker may contain a Primer pair named TaNRT2-Primer-BsmaI or a Primer pair named TaNRT 2-6A-Primer; the TaNRT-Primer-BsMI consists of two single-stranded DNAs shown as SEQ ID No.2 and SEQ ID No.3 in a sequence table; The TaNRT-6A-Primer consists of two single-stranded DNAs shown as SEQ ID No.4 and SEQ ID No.5 in a sequence table. In the above application, the kit may further comprise the restriction enzyme BsmAI. The kit may be only the TaNRT-Primer-BsmAI or the TaNRT-6A-Primer, and may also consist of the TaNRT-Primer-BsmAI and restriction enzyme BsmAI. In the application, the drought resistance of the homozygous wheat with the nucleotide A corresponding to the 1038 position of the SEQ ID No.1 in the sequence table is larger than or alternatively larger than the drought resistance of the homozygous wheat with the nucleotide G corresponding to the 1038 position of the SEQ ID No.1 in the sequence table. The invention also provides a method for detecting or assisting in detecting the drought resistance of wheat, which comprises the steps of detecting the wheat molecular marker and determining the drought resistance of wheat according to the following method: The drought resistance of the homozygous wheat with the 1038 th nucleotide corresponding to the SEQ ID No.1 in the sequence table being A is greater than or alternatively greater than the drought resistance of the homozygous wheat with the 1038 th nucleotide corresponding to the SEQ ID No.1 in the sequence table being G. In the above method, the detection of the wheat molecular marker may be performed using the substance for detecting a wheat molecular marker. In the method, the detection of the wheat molecular marker by using the TaNRT-6A-Primer can comprise the steps of carrying out PCR (polymerase chain reaction) amplification on wheat genome DNA by using the TaNRT-6A-Primer to obtain an amplification product, sequencing the amplification product, and determining a nucleotide corresponding to 1038 position of SEQ ID No.1 in a sequence table in a wheat genome. The reaction system for PCR amplification may be ddH 2 O12.2. Mu.L, 5 XPCR buffer 4.0. Mu.L, forward primer (10. Mu. Mol/L) and reverse primer (10. Mu. Mol/L) each 0.4. Mu. L, dNTPs (2.5 mmol/L) 1.6. Mu. L, transfastpfu enzyme (5U) 0.4. Mu.L, template DNA (20 ng/. Mu.L) 1. Mu.L. Both 5 XPCR buffer and transfastpfu U enzyme (5U) are available from Beijing full gold biotechnology Co. The reaction conditions of the PCR amplification can be 95 ℃ for 2min, 95 ℃ for 50s,53 ℃ for 50s,72 ℃ for 1min,35 cycles, 72 ℃ for 10min and 4 ℃ for preservation. In the method, the detection of the wheat molecular marker by using TaNRT-Primer-BsmAI can comprise the steps of carrying out PCR amplification on wheat genome DNA by using TaNRT-Primer-BsmAI to obtain an amplification product, sequencing the amplification product, and determining a nucleotide corresponding to 1038 th position of SEQ ID No.1 in a sequence table in a wheat genome. The PCR amplification reaction system may be ddH 2 O3.6. Mu.L, forward primer (10. Mu. Mol/L) and reverse primer (10. Mu. Mol/L) each 0.2. Mu.L, 2 XPCR Mix 5. Mu.L, template DNA (20 ng/. Mu.L) 1. Mu.L. Wherein 2 XPCR Mix is a product of the banker corporation, with a product number of ZT201A. The reaction conditions for PCR amplification may be 95℃for 2min, 95℃for 50s,58℃for 50s,72℃for 30s,35 cycles, 72℃for 10min, and 4℃for preservation. In the method, the detection of the wheat molecular marker by adopting TaNRT-Primer-BsmAI and BsmAI can comprise the steps of carr