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CN-122012545-A - Flavonoid glycosyltransferase and application thereof

CN122012545ACN 122012545 ACN122012545 ACN 122012545ACN-122012545-A

Abstract

The invention belongs to the technical field of biology, and particularly relates to flavonoid glycosyltransferase and application thereof. A flavonoid glycosyltransferase gene, the nucleotide sequence of which is shown in SEQ ID NO. 1. The invention focuses on the drought-resistant related genes of rice, predicts and encodes a flavonoid glycosyltransferase, provides key biochemical evidence for elucidating the molecular mechanism of the drought-resistant related genes of rice in drought response, lays a foundation for analyzing the mechanism of glycosylation modification for regulating and controlling the drought resistance of rice, and has important theoretical value and application potential for stress-resistant genetic improvement of crops through corresponding experiments.

Inventors

  • YANG QIANYING
  • PENG XIAOTONG
  • Qiu Tongxin
  • LIU ZHIHUI
  • YANG XIDONG
  • XIE QINGJUN

Assignees

  • 华南农业大学

Dates

Publication Date
20260512
Application Date
20260210

Claims (10)

  1. 1. A flavonoid glycosyltransferase gene is characterized in that the nucleotide sequence of the flavonoid glycosyltransferase gene is shown in SEQ ID NO. 1.
  2. 2. A flavonoid glycosyltransferase, which has an amino acid sequence shown in SEQ ID No. 3.
  3. 3. The application of the flavonoid glycosyltransferase gene in the synthesis of flavonoid glycoside compounds is characterized in that the nucleotide sequence of the flavonoid glycosyltransferase gene is shown as SEQ ID NO. 1.
  4. 4. The application of flavonoid glycosyltransferase in synthesizing flavonoid glycoside compounds is characterized in that the amino acid sequence of the flavonoid glycosyltransferase is shown as SEQ ID NO. 3.
  5. 5. The use according to claim 3 or 4, wherein the flavonoid glycoside compound comprises at least one of rutin, quercetin, luteolin, genistin.
  6. 6. A process for preparing flavonoid glycosides comprising the steps of: s1, obtaining flavonoid glycosyltransferase comprising an amino acid sequence shown as SEQ ID NO. 3; S2, synthesizing flavonoid glycoside compounds by using the flavonoid glycosyltransferase as a catalytic substrate in the step S1.
  7. 7. The method of preparing flavonoid glycoside compounds according to claim 6, wherein said substrate comprises flavonoid receptor and UDP-glucose, and said flavonoid glycoside compounds comprise at least one of rutin, quercetin, luteolin and genistin.
  8. 8. The method of preparing flavonoid glycoside compounds according to claim 7, wherein said flavonoid receptors comprise quercetin, luteolin and genistein.
  9. 9. A method for simultaneously detecting 7 flavonoid components in the reaction product of the flavonoid glycosyltransferase of claim 2, comprising the steps of: S1, preparing a standard solution, namely weighing rutin, quercetin, quercitrin, luteolin, genistein and genistin, dissolving with a methanol solution and dimethyl sulfoxide, fixing the volume, preparing a storage mother solution with the concentration of 10 mg/mL, and storing in a dark place at the temperature of-20 ℃; s2, preparing a sample solution, namely adding 0.1% formic acid alcohol solution into a sample to be detected, filtering, and taking filtrate as the sample solution; S3, detecting by adopting high performance liquid chromatography under the detection conditions of a chromatographic column C18, a mobile phase A of 0.1% formic acid aqueous solution, B of acetonitrile, a gradient elution program of 0-25 min 5-30% B, 25-30 min 30% B, 30-32 min 30-80% B, 32-38 min 80-5% B, 38-58 min 5% B, a flow rate of 0.4 mL.min -1 , a column temperature of 35 ℃, a sample injection volume of 20 mL and a detection wavelength of 254 nm.
  10. 10. The method of claim 9, wherein, The concentration of the methanol solution in the step S1 is 80%; The volume ratio of the methanol solution to the dimethyl sulfoxide in the step S1 is 1:1; The filtration in step S2 is carried out by using a 0.22 mm filter membrane.

Description

Flavonoid glycosyltransferase and application thereof Technical Field The invention belongs to the technical field of biology, and particularly relates to flavonoid glycosyltransferase and application thereof. Background Rice (Oryza sativa l.) provides about 80% of daily caloric intake as a staple food for more than half of the world population, especially in asia, and its safe production is critical to the assurance of food safety. However, global warming causes frequent drought, severely restricts rice yield, and causes huge economic loss. In recent years, global paddy rice is greatly reduced in yield under the influence of drought, and active oxygen accumulation induced by drought stress can cause oxidative damage to plants. While flavonoid metabolites such as flavonols are effective in scavenging active oxygen. Flavonoid metabolites, also known as flavonoids, refer to a generic term for a class of plant secondary metabolites having a 2-phenylchromone (2-phenylchromone) parent structure, which are widely present in the root, stem, leaf, flower and fruit of higher plants in either a bound (e.g., flavonoid glycoside) or free (e.g., flavonoid aglycone) form. Flavonoids are polyphenolic compounds biosynthesized by phenylalanine metabolic pathway as products of secondary metabolism of plants, and are involved in various physiological processes such as protection against environmental stress such as pathogens, herbivores and ultraviolet radiation during plant growth and development. In chemical structure, flavonoids are usually formed by connecting two benzene rings through a central three-carbon chain, and the basic skeleton can be derived into various subclasses, such as flavonols, anthocyanin, flavans and the like, and the compounds not only endow plants with color characteristics, but also have various biological activities of antioxidation, anti-inflammation, anti-tumor and the like. Glycosyltransferase has the functions of enhancing stability, bioactivity, water solubility and the like of metabolic substrates such as flavonol and the like, and can catalyze the flavonol to generate secondary metabolites with unknown functions such as flavonol glycoside and the like, thereby influencing drought resistance level of rice. Therefore, the further research on glycosyltransferase has important theoretical value and application potential for crop stress-tolerant genetic improvement. Disclosure of Invention The present invention aims to provide a flavonoid glycosyltransferase and its use. The invention focuses on the drought resistance related gene RESPONSE TO DROUGHT (RTD 1) of rice and predicts and encodes a flavonoid glycosyltransferase, and through corresponding experiments, not only provides key biochemical evidence for elucidating the molecular mechanism of the RTD1 in drought response, but also lays a foundation for analyzing the mechanism of glycosylation modification for regulating and controlling the drought resistance of the rice, and has important theoretical value and application potential for crop stress-resistant genetic improvement. In order to achieve the purpose, the invention adopts the following technical scheme that the nucleotide sequence of the flavonoid glycosyltransferase gene is shown as SEQ ID NO. 1. The invention also claims a flavonoid glycosyltransferase with an amino acid sequence shown in SEQ ID NO. 3. The invention also claims the application of a flavonoid glycosyltransferase gene in the synthesis of flavonoid glycoside compounds, and the nucleotide sequence of the flavonoid glycosyltransferase gene is shown as SEQ ID NO. 1. The invention also claims the application of the flavonoid glycosyltransferase in the synthesis of flavonoid glycoside compounds, and the amino acid sequence of the flavonoid glycosyltransferase is shown as SEQ ID NO. 3. As an embodiment of the present invention, the flavonoid glycoside compound includes at least one of rutin, quercetin, luteolin, and genistin. The invention also claims a method for preparing flavonoid glycoside compounds, comprising the steps of: s1, obtaining flavonoid glycosyltransferase comprising an amino acid sequence shown as SEQ ID NO. 3; S2, synthesizing flavonoid glycoside compounds by using the flavonoid glycosyltransferase as a catalytic substrate in the step S1. As one embodiment of the invention, the substrate comprises flavonoid receptor and UDP-glucose, and the flavonoid glycoside compound comprises at least one of rutin, quercetin, luteolin and genistin. As one embodiment of the present invention, the flavonoid receptors include quercetin, luteolin and genistein. The invention also claims a method for simultaneously detecting 7 flavonoid components in the reaction product of said flavonoid glycosyltransferase comprising the steps of: S1, preparing a standard solution, namely weighing rutin, quercetin, quercitrin, luteolin, genistein and genistin, dissolving with a methanol solution and dimethyl sulfoxide, fixing the volume, pr