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CN-122012772-A - Molecular marker of powdery mildew resistance related gene PmSESY of forest rye and application thereof

CN122012772ACN 122012772 ACN122012772 ACN 122012772ACN-122012772-A

Abstract

The invention provides a molecular marker of a forest rye powdery mildew resistance related gene PmSESY and application thereof, belongs to the technical field of plant molecular biology and genetic breeding, and relates to the molecular marker which is closely linked with a forest rye powdery mildew resistance gene PmSESY and marked as PmSESY-M1, wherein an amplification primer of the molecular marker is shown as SEQ ID No. 1 and SEQ ID No. 2, the molecular marker amplification primer can be used for PCR amplification in a 1RL chromosome fragment material containing PmSESY genes to obtain a specific product of 383 bp, and the molecular marker PmSESY-M1 has the advantages of strong specificity, high stability, good repeatability and the like, can be used for efficiently and accurately detecting and tracking a disease resistance chromosome fragment in a wheat-forest rye introgression line, provides a novel practical auxiliary selection tool for wheat powdery mildew resistance molecular breeding, and has important practical value in wheat powdery mildew resistance molecular breeding.

Inventors

  • TANG QIULIAN
  • HE HUAGANG
  • ZHU PANYING

Assignees

  • 江苏大学

Dates

Publication Date
20260512
Application Date
20260128

Claims (8)

  1. 1. The molecular marker PmSESY-M1 is characterized in that the molecular marker is used for specifically detecting the powdery mildew resistance gene PmSESY of forest rye, and the molecular marker is positioned on the 1RL chromosome of the powdery mildew resistance gene PmSESY of forest rye and is tightly linked with the powdery mildew resistance gene PmSESY of forest rye.
  2. 2. A primer for detecting powdery mildew resistance gene PmSESY or molecular marker PmSESY-M1 according to claim 1, characterized in that the nucleotide sequence of the primer is as follows: PmSESY-M1-F:5’-CTAGCTTTGATTGAGTCCATTG-3’(SEQ ID No:1), PmSESY-M1-R:5’-TGGCACGTGACAATGGCACCAT-3’(SEQ ID No:2)。
  3. 3. a product for identifying powdery mildew resistance, characterized in that it comprises a reagent or a kit, said product comprising the primer of claim 2.
  4. 4. Use of the molecular marker of claim 1, or the primer of claim 2, or the product of claim 3, comprising: (1) Detecting whether wheat carries powdery mildew resistance genes PmSESY; (2) Identifying or assisting in identifying whether forest rye, wheat or wheat-forest rye is powdery mildew resistant; (3) Powdery mildew resistant wheat breeding or auxiliary breeding.
  5. 5. A method for detecting powdery mildew resistance, the method comprising: Carrying out PCR amplification on the genomic DNA of the leaf of the material to be detected by adopting the primer of the molecular marker of claim 1 or the primer of claim 2, detecting an amplified product, and if a specific target product is obtained, obtaining the material to be detected as powdery mildew resistant material.
  6. 6. The method of claim 5, wherein the specific target product is a specific amplified fragment of 383bp in size.
  7. 7. The method of claim 5, wherein the 25. Mu.L PCR reaction system used in the PCR amplification procedure comprises 50 ng pieces of leaf genomic DNA of the material to be tested, 1 XPCR buffer, 1.5 mmol L -1 MgCl 2 ,200 mmol L -1 dNTPs, amplification primer with a final concentration of 2 mol L -1 , and 1U Taq DNA polymerase.
  8. 8. The method of claim 5, wherein the reaction sequence for PCR amplification comprises pre-denaturation at 94℃for 3 minutes, denaturation at 94℃for 20 seconds, annealing at 55℃for 30 seconds, extension at 72℃for 60 seconds, 34 cycles, extension at 72℃for 5 minutes, and preservation at 4 ℃.

Description

Molecular marker of powdery mildew resistance related gene PmSESY of forest rye and application thereof Technical Field The invention belongs to the technical field of plant molecular biology and genetic breeding, and particularly relates to a molecular marker of a powdery mildew resistance related gene PmSESY of forest rye and application thereof. Background Wheat powdery mildew is a leaf disease caused by Blumeria graminea, seriously threatens the production safety of wheat worldwide, and is the most economical and effective strategy for preventing and treating the disease by breeding and popularizing disease-resistant varieties at present. However, the disease-resistant genetic basis of the existing main cultivated wheat variety is relatively narrow, and the physiological species of pathogenic bacteria are constantly mutated, and many identified powdery mildew resistance genes gradually lose resistance. Therefore, the development of new powdery mildew resistance genes from wheat related species widens the genetic basis of wheat resistance and is important for realizing sustainable development of wheat production. Rye (Secale) species, particularly cultivated rye, are an important gene pool for wheat disease-resistant breeding. A number of powdery mildew-resistant genes, such as Pm7, pm8, pm17, pm20 and Pm56, have been successfully transferred from cultivated rye to common wheat. However, fewer powdery mildew resistance genes were mined from forest rye (SECALE SYLVESTRE). PmSESY is a novel powdery mildew resistant gene recently identified from forest rye strain SESY-01, which is located at the end of forest rye 1RL chromosome (He et al . Characterization of a new gene for resistance to wheat powdery mildew on chromosome 1RL of wild rye Secale sylvestre. Theor Appl Genet. 2021,134:887-896）,, introducing the gene into common wheat, is expected to provide new genetic resources for wheat disease-resistant breeding, however, in actual breeding, after exogenous chromosome fragments are introduced into wheat by distant hybridization, the method only relies on phenotype identification to screen disease-resistant plants, has the problems of low efficiency, long period, easy environmental interference and the like, therefore, the molecular marker closely linked with target genes and having high specificity under the background of common wheat is developed, the molecular marker for stably applying the forest rye PmSESY gene in the common wheat genetic background has not been reported at present. Disclosure of Invention Aiming at some defects existing in the prior art, the invention provides a molecular marker of a forest rye powdery mildew resistance related gene PmSESY and application thereof, wherein the molecular marker is closely linked with a forest rye powdery mildew resistance gene PmSESY and is marked as PmSESY-M1, an amplification primer of the molecular marker is shown as SEQ ID No. 1 and SEQ ID No. 2, the molecular marker amplification primer can be used for PCR amplification in a 1RL chromosome fragment material containing PmSESY genes to obtain a specific product of 383 bp, and the molecular marker PmSESY-M1 has the advantages of strong specificity, high stability, good repeatability and the like, can be used for efficiently and accurately detecting and tracking a disease resistance chromosome fragment in a wheat-forest rye introgression line, provides a novel practical auxiliary selection tool for wheat powdery mildew resistance molecular breeding, and has important practical value in wheat powdery mildew resistance molecular breeding. In order to achieve the technical purpose, the invention adopts the following technical means: The invention firstly provides a molecular marker PmSESY-M1, wherein the molecular marker is used for specifically detecting a powdery mildew resistance gene PmSESY of forest rye, and the molecular marker is positioned on a 1RL chromosome of a powdery mildew resistance gene PmSESY of forest rye and is tightly linked with a powdery mildew resistance gene PmSESY of forest rye. The invention also provides a primer for detecting powdery mildew resistance gene PmSESY or the molecular marker PmSESY-M1, wherein the nucleotide sequence of the primer is as follows: PmSESY-M1-F:5’-CTAGCTTTGATTGAGTCCATTG-3’(SEQ ID No:1), PmSESY-M1-R:5’-TGGCACGTGACAATGGCACCAT-3’(SEQ ID No:2)。 the invention also provides a product for identifying powdery mildew resistance, which comprises the primer. Preferably, the product comprises a reagent or kit. The invention also provides application of the molecular marker PmSESY-M1, or a primer or a product in detecting wheat, wherein the application comprises the following steps: (1) Detecting whether wheat carries powdery mildew resistance genes PmSESY; (2) Identifying or assisting in identifying whether forest rye, wheat or wheat-forest rye is powdery mildew resistant; (3) Powdery mildew resistant wheat breeding or auxiliary breeding. The invention also provides