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CN-121992128-A - Use of Rht-B1B for modulating flavonoid synthesis in wheat

CN121992128ACN 121992128 ACN121992128 ACN 121992128ACN-121992128-A

Abstract

The invention discloses an application of a Rht-B1B gene in regulating and controlling flavonoid compound synthesis in wheat. Use of the Rht-B1B gene as a target in screening for products that regulate flavonoid synthesis in wheat. The invention also provides application of the Rht-B1B gene, the Rht-B1B protein or related biological materials in regulating synthesis of flavonoids in wheat or preparing products regulating synthesis of flavonoids in wheat. According to the invention, the content of flavonoid compounds in wheat with the Rht-B1B gene knocked out or overexpressed is detected, so that the Rht-B1B gene has a new purpose of regulating and controlling the synthesis of the flavonoid compounds, and an important gene resource is provided for functional wheat breeding.

Inventors

  • XU SHENGCHUN
  • GOU JINYING
  • YAN YAN
  • TAO XIAOYUAN
  • WANG CHUYANG
  • YUAN LU

Assignees

  • 湘湖实验室(农业浙江省实验室)

Dates

Publication Date
20260508
Application Date
20241101

Claims (8)

  1. Use of the rht-B1B gene as a target in the screening of products regulating flavonoid synthesis in wheat.
  2. 2. The use according to claim 1 wherein the amino acid sequence encoded by the Rht-B1B gene comprises a sequence as set forth in SEQ ID No. 3; And/or the flavonoid compound is selected from one or more of luteolin, chrysoeriol, isoorientin-7-O-glucoside, isobergamotin, luteolin O-deoxyglucoside-C-glucoside, isoorientin, isovitexin-2 ' -O-glucoside, isocytidine-2 ' -O-glucoside, apigenin O-deoxyglucoside-C-glucoside, isocytidine-2 ' -O-rhamnoside, isovitexin and isocytidine; and/or, the regulation of flavonoids in wheat means increasing or decreasing the content of flavonoids in wheat by increasing or decreasing the level of Rht-B1B gene in wheat; and/or, the product for regulating the synthesis of flavonoids in wheat refers to a molecule capable of inhibiting transcription or translation of the Rht-B1B gene or capable of inhibiting expression or activity of the Rht-B1B protein so as to inhibit the synthesis of flavonoids in wheat; and/or, the product for regulating the synthesis of flavonoid compounds in wheat refers to a molecule capable of promoting transcription or translation of the Rht-B1B gene or promoting expression or activity of Rht-B1B protein so as to promote the synthesis of flavonoid compounds in wheat.
  3. Use of the Rht-B1B gene, the Rht-B1B protein or related biological material for regulating the synthesis of flavonoids in wheat or for the preparation of a product regulating the synthesis of flavonoids in wheat.
  4. Use of the Rht-B1B gene, the Rht-B1B protein or related biological material for cultivating wheat or for preparing a product for cultivating wheat.
  5. 5. The use according to claim 3 or 4, wherein the biological material is at least one of the following: a) Expression cassette, recombinant expression vector or recombinant bioengineering bacteria containing Rht-B1B gene; b) Nucleic acid molecules which inhibit the Rht-B1B gene or which reduce the content of Rht-B1B protein; c) A recombinant vector comprising the nucleic acid molecule of b); d) Recombinant bioengineering bacteria containing b) the nucleic acid molecules or bioengineering bacteria containing c) the recombinant expression vectors; the sequence of the Rht-B1B protein is shown as SEQ ID NO. 3.
  6. 6. The use according to claim 5, wherein b) the nucleic acid molecule is an sgRNA or a DNA molecule expressing said sgRNA, the target sequence of which comprises the sequences shown in SEQ ID No.6 and SEQ ID No. 7.
  7. 7. A method of modulating flavonoid synthesis in wheat, comprising one or more of the following steps: d1 Introducing the Rht-B1B gene to increase the flavonoid content; d2 Introducing biological material related to the Rht-B1B gene to increase the content of flavonoids; d3 Reducing the level of Rht-B1B protein in wheat to reduce the level of flavonoids; The amino acid sequence of the Rht-B1B is shown as SEQ ID NO. 3.
  8. 8. A method of cultivating wheat, the method comprising one or more of the following steps: D1 Introducing Rht-B1B gene to cultivate wheat with increased flavonoid content; d2 Introducing biological material related to the Rht-B1B gene to cultivate wheat with increased flavonoid content; d3 Reducing the content of Rht-B1B protein in wheat to cultivate wheat with reduced flavonoid content; The amino acid sequence of the Rht-B1B is shown as SEQ ID NO. 3.

Description

Use of Rht-B1B for modulating flavonoid synthesis in wheat Technical Field The invention belongs to the technical field of plant genetic engineering, and particularly relates to application of Rht-B1B in regulating and controlling synthesis of flavonoid compounds in wheat. Background With the increase of health consciousness, the demands for agricultural products and foods are gradually changed from 'full' to 'eating' and 'eating nutrition' and even 'eating health'. Development and utilization of agricultural products having specific functions has become a new trend in recent years for agricultural development. Wheat is taken as one of important grain crops in China, is an important source of human nutrition, is rich in seed nutrition, and has dietary therapy health care value (Avisar et al, 2020;Sharma et al, 2020;Adhikary et al and 2021) as well as wheat seedlings (if leaves and green juice). Currently, green juice is consumed by people in various forms such as tablets, freeze-dried powders and the like. However, the functional nutritional components of the wheat straw product are not clear. Therefore, the nutritional value of the wheat is further improved, the utilization range of the wheat is widened on the basis of grain stabilization and supply protection, and the method has important significance in increasing income of peasants and promoting the health level of the peasants. Flavonoids are important secondary metabolites in plants, have strong biological and pharmacological activities (Shen et al, 2022), and have multiple effects of antioxidation, antibiosis and antiphlogosis, and enhancing human immunity, etc., and thus have become one of targets for improving crop nutrition quality (Tian et al, 2022; chen Jiehe Chen Wei, 2023). Screening and creation of high-flavonoid germplasm has been carried out in crops such as rice and wheat (Chen et al 2020; wang Yanxun et al 2023). Related studies have shown that the regulation of key genes of the flavonoid synthesis pathway by molecular genetic means helps to further increase the flavonoid content in foods and to increase their health efficacy (Tao et al, 2022). Therefore, the molecular mechanism and genetic basis affecting the flavonoid content are systematically analyzed, so that the method is not only expected to meet the increasing demands of consumers on nutrition quality, but also can provide genes, germplasm resources and theoretical basis for the development and utilization of functional agricultural products. Flavonoids are a generic term for a number of substances with similar structures in plants, whose synthesis starts in the phenylpropane pathway (Yonekura-Sakakibara et al, 2019). The flavonoid content is affected by enzymes involved in the flavonoid synthesis pathway and by various regulatory factors that control the expression levels of synthetic genes in the phenylpropane metabolic pathway (Dong and Lin,2021; lam et al, 2023). Among them, the DELLA protein belongs to the GRAS family specific to plants, is the main negative regulator of gibberellin (Gibberellic acid, GA) signaling pathway, and can integrate multiple signaling pathways through direct interaction with many transcription and co-transcription factors, and the regulation of DELLA protein in phenylpropane metabolic pathway has been reported in early studies. For example, in anthocyanin biosynthesis, GA signaling has been demonstrated to inhibit its biosynthesis by the DELLA protein RGA. The expression of anthocyanin-specific genes is regulated by conserved MBW complexes (consisting of MYB, bHLH and WD40 subunits), but MBW activity is inhibited by MYBL2 and JAZ proteins. RGA, in turn, binds to MYBL2 and JAZ proteins, releasing the bHLH/MYB subunit, which forms an active MBW complex, which in turn activates the anthocyanin biosynthetic pathway (Xie et al, 2016). In addition, RGA is also capable of interacting with transcription factors MYB12/MYB111 in the flavonol synthesis pathway and promoting binding of MYB12 to the downstream target gene FLS1/CHS promoter, thereby activating expression of flavonol synthesis pathway genes (Tan et al, 2019). Thus, DELLA proteins can be seen to have important regulatory roles in flavonoid biosynthesis. Wheat (Triticum aestivum l.) is an important food crop. The Rht-1 dwarf gene codes DELLA protein and is an important genetic basis for wheat semi-dwarf breeding. In hexaploid wheat, DELLA proteins are encoded by 3 homologous Rht-1 genes, rht-A1a, rht-B1a and Rht-D1a, respectively. Allelic variation Rht-B1B was generated by single base mutation of Rht-B1a near the DELLA region. Mutation of Rht-B1B results in premature termination of translation of Rht-1 encoded protein, followed by translation restart encoding to produce N-terminally truncated DELLA protein Rht-B1B. Rht-B1B is insensitive to gibberellin signals, resulting in semi-dwarfing of plants, but has not been reported in related studies on the regulation of wheat flavonoid content. Disclosure o