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CN-122012779-A - InDel marker related to fat content of millet seeds and application thereof

CN122012779ACN 122012779 ACN122012779 ACN 122012779ACN-122012779-A

Abstract

The invention discloses an InDel marker related to the fat content of millet grains and application thereof, and belongs to the technical field of genetic engineering. The InDel marker provided by the invention is a 792 bp insertion variation positioned on chromosome 5 of the millet genome, and the nucleotide sequence of the InDel marker is shown as SEQ ID NO. 1. The invention develops a primer group and a detection method for detecting the InDel marker. Experimental results show that the primer group and the detection method provided by the invention can be used for rapidly and accurately identifying the fat content of the millet seeds, and the detection method provided by the invention also has the advantages of strong applicability, stable effect, simplicity and convenience in operation, low detection cost and the like. The invention provides a new molecular marker for the auxiliary selective breeding of the high-quality properties of the millet grains and a new technical means for cultivating or screening high/low-fat millet varieties.

Inventors

  • ZHANG BIN
  • MA QI
  • WAN XIN
  • HAN YUANHUAI

Assignees

  • 山西农业大学

Dates

Publication Date
20260512
Application Date
20260210

Claims (10)

  1. 1. An InDel marker related to the fat content of millet grains, which is characterized in that the nucleotide sequence of the InDel molecular marker is shown as SEQ ID NO. 1.
  2. 2. A primer group for amplifying the InDel marker according to claim 1, which comprises an upstream primer with a nucleotide sequence shown as SEQ ID NO.2 and a downstream primer with a nucleotide sequence shown as SEQ ID NO. 3.
  3. 3. Use of the primer set of claim 2 for preparing a product for identifying fat content of millet kernels.
  4. 4. The use according to claim 3, wherein the product is a kit.
  5. 5. A product for identifying the fat content of millet kernels, comprising the primer set of claim 2.
  6. 6. Use of a primer set according to claim 2 or a product according to claim 5 for identifying the fat content of millet kernels.
  7. 7. A method for identifying the fat content of millet kernels, comprising the steps of: Extracting the DNA of the millet to be detected, and carrying out PCR amplification by using the primer set of claim 2 or the product of claim 5 to obtain an amplification product; electrophoresis, staining and developing are carried out on the PCR amplification product, and a band is interpreted; The interpretation strip is: if 1110 bp single bands are amplified, the millet to be detected is the millet with high grain fat content; If a single band of 318 bp is amplified, the millet to be detected is a millet with low grain fat content.
  8. 8. Use of a primer set according to claim 2 or a product according to claim 5 in breeding of grain fat content of millet.
  9. 9. The breeding method of the millet with high grain fat content is characterized by comprising the following steps of: Extracting the DNA of the millet to be detected, and carrying out PCR amplification by using the primer set of claim 2 or the product of claim 5 to obtain an amplification product; electrophoresis, staining and developing are carried out on the PCR amplification product, and a band is interpreted; The interpretation strip is: if 1110 bp single bands are amplified, retaining the millet to be tested for breeding.
  10. 10. The breeding method of the millet with low fat content of the grains is characterized by comprising the following steps of: Extracting the DNA of the millet to be detected, and carrying out PCR amplification by using the primer set of claim 2 or the product of claim 5 to obtain an amplification product; electrophoresis, staining and developing are carried out on the PCR amplification product, and a band is interpreted; The interpretation strip is: if a single band of 318 bp is amplified, retaining the millet to be tested for breeding.

Description

InDel marker related to fat content of millet seeds and application thereof Technical Field The invention relates to the technical field of genetic engineering, in particular to an InDel marker related to the fat content of millet grains and application thereof. Background Millet (SETARIA ITALICA) is an important coarse cereal crop, has the characteristics of short growth period, drought resistance and barren resistance, is rich in nutrition, contains multiple nutritional ingredients such as multiple amino acids, fatty acids, vitamins and the like, has high digestibility, improves human immunity, prevents diseases and other important dietary therapy functions, and is popular with consumers. Vegetable fat is mainly stored in oil crop seeds, is a main energy source in germination, and also provides important fatty acid intake sources in foods for human beings, and is an important biological energy source applied to chemical production. The fatty acid content is more than 90% of the total fat, so the nature and the nutritional value of the fat mainly depend on the composition of the fatty acid, the fatty acid can be divided into saturated fatty acid and unsaturated fatty acid, the excessive intake of the saturated fatty acid by human body can cause diseases such as fatty liver, hyperlipidemia, arteriosclerosis and the like, and the unsaturated fatty acid is a well-known beneficial component, and the intake of the polyunsaturated fatty acid has indispensable functions in the aspects of vision maintenance, anti-inflammatory response regulation and the like. The average content of millet fat is 4.05%, and the ratio of unsaturated fatty acid is 80%, wherein the millet fat is rich in linoleic acid necessary for human body, and has effects of dilating blood vessel and preventing thrombosis. The fat content and composition of the millet kernel not only determine the nutritional quality of the millet kernel, but also have potential correlation with the agronomic traits of crops. Researches show that the fat metabolism process is possibly involved in regulating and controlling the yield-related characters such as weight, plumpness and the like of grains, the rule provides theoretical reference for the cooperative improvement of the yield and the quality of the grains, and the importance of accurately regulating and controlling the fat content of the grains to breeding work is also highlighted. At present, molecular breeding research of millet is focused on the directions of stress resistance, leaf color and the like, molecular marker development relative hysteresis aiming at the fat content of seeds is not reported and applied to breeding practice, and InDel markers which are stably linked with the fat content and fatty acid composition of the seeds of the millet are not yet reported. Therefore, the InDel marker closely linked with the fat content of the millet seeds is mined, a rapid and accurate molecular detection method is established, the early screening and directional improvement of the fat content characters of the millet are realized, the breeding period is shortened, the breeding efficiency is improved, and the method has important practical significance and application value for promoting the cultivation of high-quality millet varieties and promoting the high-quality development of the millet industry. Disclosure of Invention The invention aims to provide an InDel marker related to the fat content of millet seeds and application thereof, so as to solve the problems in the prior art, and provides an InDel molecular marker for detecting the fat content of the millet seeds. In order to achieve the above object, the present invention provides the following solutions: the invention provides an InDel marker related to the fat content of millet grains, and the nucleotide sequence of the InDel molecular marker is shown as SEQ ID NO. 1. The invention also provides a primer group for amplifying the InDel marker, which comprises an upstream primer with a nucleotide sequence shown as SEQ ID NO.2 and a downstream primer with a nucleotide sequence shown as SEQ ID NO. 3. The invention also provides application of the primer group in preparation of products for identifying fat content of millet grains. Further, the product is a kit. The invention also provides a product for identifying the fat content of the millet grains, which comprises the primer group. The invention also provides an application of the primer group or the product in distinguishing the fat content of the millet seeds. The invention also provides a method for identifying the fat content of the millet grains, which comprises the following steps: Extracting the DNA of the millet to be detected, and carrying out PCR amplification by using the primer group or the product to obtain an amplified product; electrophoresis, staining and developing are carried out on the PCR amplification product, and a band is interpreted; The interpretation strip is: if 1110 bp si