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CN-121974991-A - Application of ZmMADS1 gene in regulation and control of folic acid content in corn

CN121974991ACN 121974991 ACN121974991 ACN 121974991ACN-121974991-A

Abstract

The invention relates to the technical field of genetic engineering, in particular to application of ZmMADS1 gene in regulating and controlling corn folic acid content. Experiments show that the ZmMADS1 gene negatively regulates the activity of an ADCS gene promoter, so that the content of folic acid in corn is regulated, the content of folic acid in corn can be improved by knocking out the ZmMADS1 gene, and a new variety of high-folic acid corn can be cultivated.

Inventors

  • SONG LILI
  • LV BEIBEI
  • WU GUOGAN
  • WANG CUI
  • SUN YU

Assignees

  • 上海市农业科学院

Dates

Publication Date
20260505
Application Date
20250924

Claims (10)

  1. The application of ZmMADS1 gene in regulating and controlling corn folic acid content, wherein the amino acid sequence of protein coded by the ZmMADS1 gene is shown as SEQ ID NO. 25.
  2. 2. The method according to claim 1, wherein the controlling comprises decreasing the expression level of ZmMADS1 gene and increasing the folic acid content of the corn.
  3. 3. The use according to claim 2, wherein the method for reducing the expression level of ZmMADS1 gene comprises gene editing.
  4. 4. The use according to any one of claims 1 to 3, wherein the nucleotide sequence of the ZmMADS1 gene is shown in SEQ ID No. 23.
  5. 5. The use according to claim 1 or 2, wherein said modulation of corn folate content comprises modulation of folate content in corn kernels.
  6. 6. A gene editing vector for improving the folic acid content of corn is characterized in that a target sequence of the gene editing vector comprises target sequences with nucleotide sequences shown as SEQ ID NO. 3-SEQ ID NO.5, and an amino acid sequence of a protein coded by a ZmMADS1 gene is shown as SEQ ID NO. 25.
  7. 7. The gene editing vector according to claim 6, wherein the gene editing vector comprises an editing vector and sgRNA, and the nucleotide sequence of the sgRNA is shown as SEQ ID NO. 6-SEQ ID NO. 11.
  8. 8. The gene editing vector of claim 7, wherein the editing vector comprises a 143 vector.
  9. 9. An engineering bacterium for increasing the folic acid content of corn, which is characterized by comprising agrobacterium and the gene editing vector of any one of claims 6-8 introduced into agrobacterium.
  10. 10. The use of the gene editing vector of any one of claims 6 to 8 or the engineering bacterium of claim 9 for increasing the folic acid content of corn.

Description

Application of ZmMADS1 gene in regulation and control of folic acid content in corn Technical Field The invention relates to the technical field of genetic engineering, in particular to application of ZmMADS1 gene in regulating and controlling corn folic acid content. Background Folic acid, also known as vitamin B9, is a water-soluble B-group vitamin that is critical to the growth and development of animals and plants. It includes Tetrahydrofolate (THF) and its derivatives, which participate in C1 transfer reactions, playing an important role in the biosynthesis of purines, thymidylates, DNA, amino acids and proteins and in the methyl cycle. Folate deficiency is associated with a variety of health problems including neural tube defects, cardiovascular disease, and the like. Since humans cannot synthesize folic acid by themselves, it must be ingested through food or supplements. The problem of folate deficiency is prevalent worldwide, particularly in areas where folate content is low in major food crops such as rice, wheat and maize. At present, medicament supplementation and crop folic acid strengthening are two effective ways for solving the problem of folic acid deficiency, and especially crop folic acid strengthening is particularly important for remote and poor areas. By increasing the folic acid content of crops through biological enhancement, not only can the nutritional status be improved, but also the problem of global folic acid deficiency can be solved as a strategy with high cost efficiency. Mutations in the methylene tetrahydrofolate reductase (MTHFR) gene may lead to folate deficiency, affect the absorption and metabolism of folic acid tablets, and increase the risk of potential diseases such as colon cancer. Thus, increasing folate levels by bioaugmentation of staple food crops is considered a green and healthy approach to combat folate deficiency. The study on the folic acid biosynthesis pathway is clear. Increased folate levels can be achieved by increasing gene transcription levels of key enzymes in the folate synthesis branch, such as GCHI (GTP cyclohydrolase) and ADCS (amino deoxychorismate synthase), two important enzymes in the folate synthesis pathway. In wheat, gmGCHI/GmADCS of soybean source is overexpressed, so that the folic acid content can be increased by 62% -230%. In raw vegetables and beans, GCHI of Arabidopsis sources are overexpressed, so that the folic acid content is respectively increased by 2.1-8.5 times and 3.0 times. Studies have shown that over-expression of GCHI and ADCS simultaneously has a higher effect on increasing folate levels than over-expression of one of the genes alone. However, the relationship between the expression pattern of the key enzyme for folate synthesis and the folic acid content in different plants is greatly different, which indicates that a complex regulation mechanism exists in the folic acid biosynthesis pathway, and specific in-vivo molecular mechanism exploration is required for specific crops in order to more effectively develop crop folic acid bioaugmentation research. Currently, functional research and application of key genes for regulating corn folic acid biosynthesis are lacking. Disclosure of Invention In order to solve the problems, the invention provides application of ZmMADS1 gene in regulating and controlling the folic acid content of corn. The ZmMADS1 gene can be used for negatively regulating and controlling the folic acid content of corn, and the folic acid content of corn can be improved by knocking out or knocking down the expression quantity of the ZmMADS1 gene, so that a new variety of high-folic acid corn is cultivated. In order to achieve the above object, the present invention provides the following technical solutions: The invention provides application of ZmMADS1 gene in regulating and controlling corn folic acid content, wherein the amino acid sequence of protein coded by the ZmMADS1 gene is shown as SEQ ID NO. 25. Preferably, the regulation comprises reducing the expression level of ZmMADS1 gene and increasing the folic acid content of corn. Preferably, the method for reducing the expression level of ZmMADS1 gene comprises gene editing. Preferably, the nucleotide sequence of the ZmMADS1 gene is shown as SEQ ID NO. 23. Preferably, the controlling the folate content of the corn comprises controlling the folate content of the corn kernel. The invention provides a gene editing vector for improving corn folic acid content, wherein a target sequence of the gene editing vector comprises target sequences with nucleotide sequences shown as SEQ ID NO. 3-SEQ ID NO.5, and an amino acid sequence of protein coded by ZmMADS1 gene is shown as SEQ ID NO. 25. Preferably, the gene editing vector comprises an editing vector and sgRNA, and the nucleotide sequence of the sgRNA is shown as SEQ ID NO. 6-SEQ ID NO. 11. Preferably, the editing carrier includes 143 carrier. The invention provides an engineering bacterium for improving the fo