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CN-121992019-A - SiMYB113 protein 113 and application of coding gene thereof in regulation and control of plant tocopherol synthesis

CN121992019ACN 121992019 ACN121992019 ACN 121992019ACN-121992019-A

Abstract

The invention discloses SiMYB113,113 protein and application of a coding gene thereof in regulating and controlling synthesis of plant tocopherol, belonging to the technical field of plant genetic engineering. The amino acid sequence of SiMYB113,113 protein is shown as SEQ ID NO.1, the invention reports the function and application of sesame SiMYB113,113 gene in tocopherol synthesis for the first time, and further constructs SiMYB gene overexpression vector to make it overexpress in sesame hairy root, verifies SiMYB in transgenic sesame hairy root can induce tocopherol synthesis, verifies SiMYB113,113 gene has the function of positive regulation of tocopherol synthesis, further improves the tocopherol content in sesame, and has important significance for analyzing sesame tocopherol content genetic regulation mechanism, cultivating new high-tocopherol sesame variety and improving sesame nutrition quality.

Inventors

  • WANG LINHAI
  • YOU JUN
  • Luo Zishu
  • ZHOU RONG

Assignees

  • 中国农业科学院油料作物研究所

Dates

Publication Date
20260508
Application Date
20260210

Claims (10)

  1. 1. Use of SiMYB protein 113 for regulating plant tocopherol synthesis, wherein the SiMYB protein is selected from any one of the following: a1 Protein with the amino acid sequence shown as SEQ ID NO. 1; A2 A protein having the same function obtained by substitution, deletion or addition of one or more amino acids as compared with the amino acid sequence defined in A1); a3 A protein having the same function obtained by ligating a tag to the N-terminal and/or C-terminal of A1) or A2).
  2. 2. Use of a nucleic acid molecule encoding the SiMYB113,113 protein of claim 1 for regulating plant tocopherol synthesis.
  3. 3. The use according to claim 2, wherein the nucleic acid molecule is selected from any one of the following: B1 A nucleic acid molecule with a nucleotide sequence shown as SEQ ID NO. 2; B2 A nucleic acid molecule which hybridizes to a nucleic acid molecule as defined in B1) and which codes for a SiMYB protein according to claim 1; B3 A nucleic acid molecule which has more than 90% sequence identity to a nucleic acid molecule as defined in B1) or B2) and which encodes a SiMYB protein according to claim 1.
  4. 4. Use of a recombinant vector comprising the nucleic acid molecule of any one of claims 2-3 for regulating tocopherol synthesis in plants.
  5. 5. Use of a recombinant cell comprising the nucleic acid molecule of any one of claims 2-3 or a recombinant cell comprising the recombinant vector of claim 4 for regulating tocopherol synthesis in a plant.
  6. 6. A method of growing a plant having an increased tocopherol content, said method comprising increasing the amount and/or activity of the SiMYB113,113 protein of claim 1 in a plant of interest to produce a plant having an increased tocopherol content compared to said plant of interest.
  7. 7. The method according to claim 6, wherein the increase in the amount and/or activity of the SiMYB protein of claim 1 in the plant of interest is achieved by increasing the expression level of the SiMYB protein-encoding gene in the plant of interest.
  8. 8. The method of claim 7, wherein the increasing the expression level of the SiMYB113,113 protein-encoding gene in the plant of interest is achieved by at least one of: C1 Increasing the copy number of the gene encoding SiMYB113,113 protein; c2 Placing SiMYB-113-protein coding gene under the drive of strong promoter to make expression; c3 Increasing the regulatory elements of the gene encoding SiMYB113,113 protein to over-express, wherein the regulatory elements comprise an enhancer element, an element that enhances mRNA stability, an element that enhances translational efficiency, and/or an element that enhances protein secretion; C4 Codon optimization of the gene encoding SiMYB113,113 protein.
  9. 9. The method according to claim 6, characterized in that said method for growing plants with increased tocopherol content comprises in particular the following steps: 1) Constructing a recombinant vector comprising a nucleic acid molecule encoding SiMYB113,113 proteins; 2) Introducing the recombinant vector into a plant of interest; 3) And screening and identifying to obtain the plant with increased tocopherol content.
  10. 10. The use according to any one of claims 1 to 5 or the method according to any one of claims 6 to 9, wherein the plant comprises sesame.

Description

SiMYB113 protein 113 and application of coding gene thereof in regulation and control of plant tocopherol synthesis Technical Field The invention belongs to the technical field of plant genetic engineering, and particularly relates to SiMYB protein and application of a coding gene thereof in regulating and controlling synthesis of plant tocopherol. Background Tocopherols, also known as vitamin E, were found in 1922. Tocopherols are a generic term for tocopherols and tocotrienols, and naturally occurring tocopherols include the 8 types of alpha, beta, gamma, delta-tocopherols and alpha, beta, gamma, delta-tocotrienols, mainly hydrophobic tail saturation and the number and position differences of methyl groups on the aromatic ring. Tocopherols are fat-soluble vitamins which are indispensable in humans and animals but cannot be autonomously synthesized, and are well known for their antioxidant function. As a natural antioxidant, the tocopherol can effectively relieve the non-enzymatic oxidation damage of free radicals or active oxygen to cells and lipid, inhibit lipid peroxidation and protect biological membranes. There have been many studies showing various potential beneficial effects of tocopherol on human health, such as anticancer, anti-atherosclerosis, neuroprotection, and inhibition of chronic diseases such as Alzheimer's disease. Thus increasing the tocopherol content in plants, especially in crops, is of great importance for promoting human health. The synthetic pathway of tocopherol in plants has been gradually clarified, but less reports on tocopherol synthesis regulatory genes are reported. Sesame (Sesamum indicum ml.) is an annual herbaceous oil crop of sesames of sesamiaceae, widely planted in asia, africa tropical and subtropical regions, and distributed in more than 70 countries with a global planting area exceeding 2.0 hundred million mu. Sesame planting history in China is long, the sesame is planted in all regions of China, main production areas are concentrated in Huang-Huai, jiang-Huai and Yangtze river basin (Henan, hubei, anhui, jiang-xi and other places), and the planting area and yield are in the front of the world. The sesame can be used for oil pressing or direct eating, can also be used as spice, medicine and chemical raw materials, and has wide application. The multiple uses of sesame are indistinguishable from their specific nutritional quality. Sesame is not only rich in unsaturated fatty acid, various minerals and proteins, but also rich in lignans, tocopherols and other antioxidant components. The sesame seeds have higher tocopherol content, and are one of ideal food sources for supplementing tocopherol. However, no report is made on the role of SiMYB gene in regulating tocopherol synthesis in plants. Disclosure of Invention The invention aims to provide SiMYB protein 113 and application of a coding gene thereof in regulating and controlling synthesis of plant tocopherols. The SiMYB protein is used for solving the problem that SiMYB protein and encoding genes thereof for efficiently regulating and controlling the synthesis of plant tocopherols are lacked in the prior art. In a first aspect, the invention provides application of SiMYB113,113 protein in regulating plant tocopherol synthesis, wherein SiMYB protein is selected from any one of A1) protein with an amino acid sequence shown as SEQ ID NO.1, A2) protein with the same function obtained by substituting, deleting or adding one or more amino acids compared with the amino acid sequence defined by A1), and A3) protein with the same function obtained by connecting a label at the N end and/or the C end of A1) or A2). The SiMYB113,113 protein provided by the present invention may be a naturally occurring, recombinant or synthetic active polypeptide, which may be a naturally purified product, a chemically synthesized product, or a product produced from a prokaryotic host (e.g., E.coli) or a eukaryotic host (e.g., yeast, higher plants) using recombinant techniques. In the present invention A3), the connection may be directly connected by peptide bond or connected by linker, and the connection method is conventional in the art. Wherein the tag includes, but is not limited to, GST (glutathione-sulfhydryl transferase) tag protein, trx (thioredoxin) tag protein, his tag protein (His-tag), flag tag protein, lacZ tag protein, GFP (green fluorescent protein), sfGFP (hyper-folded green fluorescent protein), HA tag (hemagglutinin tag). One skilled in the art can select an appropriate tag protein according to the actual use requirements. The use of the tag does not alter the function of the protein of interest (SiMYB protein), which is aimed at isolation, purification, detection or tracking. The tag may be separated from the protein of interest (SiMYB protein) by chemical cleavage or enzymatic methods known in the art (e.g., introducing a proteolytic cleavage site to cleave the tag using TEV protease). In some embodiments, the use is accomp