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CN-121992146-A - Molecular marker related to oat lodging index and application thereof

CN121992146ACN 121992146 ACN121992146 ACN 121992146ACN-121992146-A

Abstract

The invention discloses a molecular marker related to an oat lodging index and application thereof, and belongs to the technical field of molecular biology. The invention provides a molecular marker related to oat lodging indexes, which is characterized in that specific information of the molecular marker is chromosome chr7D, position 104407598 and SNP typing G/T, and aims to solve the problems of time consumption, labor consumption, low accuracy and the like of a conventional single plant selection method for breeding new lodging-resistant oat varieties in common fields. The invention also designs a primer combination aiming at the molecular marker, and the primer combination can directly and specifically distinguish and detect the G or T base of the SNP mutation site. The method has good application value, can realize the pre-selection and molecular auxiliary breeding of the lodging index character of the oat, and has important theoretical and practical significance for accelerating the genetic improvement process of breeding of suitable lodging index materials and improving the selection efficiency.

Inventors

  • ZHANG TIEJUN
  • KANG JUNMEI
  • LI XIAO
  • LIU HUANCHENG
  • ZHAO LIJUN

Assignees

  • 中国农业科学院北京畜牧兽医研究所

Dates

Publication Date
20260508
Application Date
20260408

Claims (10)

  1. 1. A molecular marker related to an oat lodging index is characterized by comprising the specific information of chromosome chr7D, position 104407598 and SNP typing G/T.
  2. 2. The molecular marker according to claim 1, wherein the nucleotide sequence of the molecular marker is shown as SEQ ID No.1 or SEQ ID No. 2.
  3. 3. The primer combination is characterized by being used for amplifying the molecular marker of any one of claims 1-2, and the nucleotide sequence is shown as SEQ ID No. 3-SEQ ID No. 4.
  4. 4. A kit is characterized in that, comprising the primer combination of claim 3.
  5. 5. A molecular marker chip comprising the primer combination of claim 3.
  6. 6. Use of the molecular marker according to any one of claims 1-2, the primer combination according to claim 3, the kit according to claim 4 and/or the molecular marker chip according to claim 5 in any one of the following optional: a. Predicting an oat lodging index; b. Identifying and screening oat with different lodging indexes; c. molecular markers assist in breeding oat materials with different lodging indexes; d. Breeding oat materials with different lodging indexes; e. preparing oat breeding products with different lodging indexes; And if the parting result is GG, the oat material with high lodging index is obtained, and if the parting result is GT, the oat material with low lodging index is obtained.
  7. 7. A method for screening oat materials with different lodging indexes or predicting the lodging indexes of the oat comprises the following steps of extracting genome DNA of the oat materials to be tested, sequencing amplified products by adopting primer combination amplified molecular markers described by SEQ ID No. 3-SEQ ID No.4, and screening in a parting mode, wherein the standard of the parting screening is that if a parting result is GG, the material with high lodging index is obtained, and if the parting result is GT, the material with low lodging index is obtained.
  8. 8. The method of claim 7, wherein the amplification procedure is 94℃for 15min, 95℃for 20sec,65 to 56℃for 60sec,10 cycles, each cycle of annealing extension temperature decrease of 0.8℃and 94℃for 20sec,57℃for 60sec,30 cycles.
  9. 9. A method for cultivating oat with different lodging indexes is characterized by comprising the following steps of identifying oat genotypes by the method of claim 7, screening target single plants according to genotypes, and breeding the screened target single plants to obtain oat with different lodging indexes.
  10. 10. The method according to claim 9, wherein the screening method of the target individual is to screen an individual with a genotype GT if an oat with a low lodging index is to be obtained, and to screen an individual with a genotype GG if an oat with a high lodging index is to be obtained.

Description

Molecular marker related to oat lodging index and application thereof Technical Field The invention belongs to the technical field of molecular biology, and particularly relates to a molecular marker related to an oat lodging index and application thereof. Background Oat is a dual-purpose crop for annual grains and feeds of the genus Avena of the family Gramineae. The lodging resistance of oat is determined by a plurality of factors, wherein the lodging index is the comprehensive expression of plant weight, gravity center height and mechanical strength of the stalk, and is an important reference basis for judging the lodging resistance of oat plants. Studies have shown that lower lodging indices represent stronger lodging resistance and higher values indicate weaker lodging resistance. At present, oat breeding in China is still in a conventional breeding stage, and excavation and identification of important indexes such as lodging indexes are not deep enough. Through researching the related genes of the lodging index and the regulation and control mechanism thereof, the molecular marker closely linked with the lodging index can be developed and used for molecular marker assisted breeding, and the breeding efficiency and accuracy are improved. The selection of the new variety of oat with low lodging index is based on the evaluation and selection of lodging index of the offspring of the traditional variety, and the method is time-consuming, labor-consuming and low in accuracy. The development of specific molecular markers for auxiliary selection is the best method for improving the selection efficiency by utilizing the base difference of target genes. Competitive allele-specific PCR (Kompetitive Allele-SPECIFIC PCR, KASP) molecular markers are a novel SNP typing method based on allele-specific amplification (Amplification Refractory Mutation System, ARMS) and high-sensitivity fluorescent detection. The principle is that a forward primer and a reverse primer are designed aiming at an allele SNP locus, and each pair of forward and reverse primers has a specific sequence and can be combined with different fluorescent markers. The PCR amplification of the DNA of the sample to be tested with forward and reverse primers with sequences binding to different fluorescence can be reflected by different fluorescent signals (HeC L, et al SNP genotyping: the KASP assay. Methods mol biol,2014,1145: 75-86). Therefore, the KASP marker closely linked with the oat lodging index is developed and used for early selection of breeding, is important to reduce the breeding workload and accelerate the oat lodging-resistant breeding, and has obvious economic benefit. Disclosure of Invention Aiming at the problems of time consumption, labor consumption and low accuracy of the traditional new variety breeding method of the lodging-resistant oat, the invention provides a molecular marker related to the lodging index of the oat, wherein specific information of the molecular marker is chromosome chr7D, position 104407598 and SNP typing G/T. The invention also designs a primer combination aiming at the molecular marker, and the primer combination can directly and specifically distinguish and detect the G or T base of the SNP mutation site. The method has good application value, can realize the pre-selection and molecular auxiliary breeding of the lodging index character of the oat, and has important theoretical and practical significance for accelerating the genetic improvement process of breeding of suitable lodging index materials and improving the selection efficiency. In order to achieve the technical purpose, the invention adopts the following technical scheme: The invention aims to provide a molecular marker related to an oat lodging index, and specific information of the molecular marker is chromosome chr7D, position 104407598 and SNP typing G/T. Further, the nucleotide sequence of the molecular marker is shown as SEQ ID No.1 or SEQ ID No. 2. The second object of the invention is to provide a primer combination for amplifying the molecular marker, wherein the nucleotide sequence is shown as SEQ ID No. 3-SEQ ID No. 4. The third object of the present invention is to provide a kit comprising the primer combination. The fourth object of the present invention is to provide a molecular marker chip, which comprises the primer combination. It is a fifth object of the present invention to provide the use of said molecular markers, said primer combinations, said kits and/or said molecular marker chips in any one of the following optional items: a. Predicting an oat lodging index; b. Identifying and screening oat with different lodging indexes; c. molecular markers assist in breeding oat materials with different lodging indexes; d. Breeding oat materials with different lodging indexes; e. preparing oat breeding products with different lodging indexes; And if the parting result is GG, the oat material with high lodging index is obtained, and if t